Mycoplasmas Properties and Their Role in Autoimmune Diseases

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.isolates were referred to as Pleuropneumonia-Like Organisms(PPLO), now known as Mycoplasmas in the broader Mollecutesfamily.The terms PPLO and wall-less bacteria L-forms were usedinterchangeably until 1962 when a viral-like agent causingatypical pneumonia but sensitive to tetracycline and cultured in acell-free media was found to be a new human mycoplasma strain(M. pneumoniae). Because of the increasing number of strains,®PPLO were classified under bacterial Mollecutes with theSan Marcos University College of Veterinarian Medicine mycoplasmatacae family and subspecies mycoplasma andLima, Peru Lectures 8/99ureaplasma. Our modern day culture techniques have improvedby Harold W. Clark, Ph.D.over the early technique of implanting a cellophane bag with theMycoplasma Research Institute, PO Box 640040, Beverly Hills, specimen in a rabbit’s abdomen. The difficulties in isolatingFL 34464-0040; E-mail: www.hwcmri@hitter.net mycoplasmas from inflamed tissues still remains.All rights reserved by the The Roger Wyburn-Mason and Jack M.BlountSince their first isolation, a major impediment to mycoplasmaFoundation for Eradication of Rheumatoid Diseaseresearch and laboratory diagnosis of infection has been theAKA The Arthritis Trust of America ®difficulty of isolating mycoplasmas from the host tissues. To7376 Walker Road, Fairview, TN 37062overcome this problem complex cell-free media are used for bothwww.arthritistrust.orgisolation and cultivation. The media is usually based on beef heartinfusion, peptone digest, yeast extract, serum and varioussupplements. The continued use of these complex undefinedmedia components has limited the precise definition ofmycoplasma’s metabolic pathways, genetic analysis, preparationof mycoplasma antigens free of media components and specificmonoclonal antiserum free of endogenous antibodies. The abilityto detect and identify mycoplasmas in tissues using the geneticPolymerase Chain Reaction (PCR) technique should be measuredin relation to the host’s immunologic and other responses to aninfection. In addition there is some concern about the adaptation ofnew mycoplasma isolates being changed when grown in artificialmedia and subjected to viruses.The first question we should ask and define is “What areHarold W. Clark, Ph.D.Mycoplasmas?” And what are their connections with humanarthritis especially the chronic rheumatic diseases of unknownLecture I: Mycoplasmas Properties andorigin. If mycoplasmas are now known to cause arthritis in animalstheir Role in Autoimmune Diseasesas well as acute and chronic disease in human lungs and theThank you for inviting me to lecture on Mycoplasma Arthritisgenitourinary tract their location in human joints could also be thein humans.irritant causing inflammation. Thus we have mixed tissue arthritisToday I will review “Mycoplasmas Properties and their Rolewith a combination of symptoms. Mixed arthritis is furtherin Autoimmune Diseases,” by Harold W. Clark, PhDcomplicated by adding another vague term “RheumatoidIn 1952 when I joined Dr. Thomas McPherson Brown, chiefArthritis” (RA). According to the U.S. Arthritis Foundation, theof medicine at the George Washington Univ. Medical School inwhole group of Rheumatic Diseases now constitutes 171 types ofWashington, D.C, as Research Biochemist, the chief of medicine atarthritis with associated diagnoses and mixtures of symptoms.the University of Rochester in New York told me that Dr. BrownThese include mycoplasma arthritis, connective tissue diseases,has a theory that some atypical microbe was the cause ofand mixed connective tissue diseases. RA was initially calledRheumatoid Arthritis that should be treated with antibiotics.Chronic Infectious Arthritis, when it was first thought to be causedUnknown to me, the year before, 1951, Dr. Brown and staff had justby some atypical viral-like agent and is now referred to aspublished and reported at the International Arthritis meeting theirAutoimmune disease, Immune Complex disorders, and Collagenrecent findings entitled “A Study of the Antigen-AntibodyVascular disorders. The diagnostic terminology is more confusingMechanism in Rheumatoid Diseases”. This report emphasized thewhen based on the symptoms, the tissue targeted, the diseaseantibiotic treatment of rheumatoid diseases based on the suspectedmechanisms, and the microbial causes. This terminology includesmycoplasma role in the inflammatory immune complex thatseptic, reactive or infectious, bacterial, viral, Lyme, andshould be blocked by immunosuppresing agents.mycoplasma arthritis in humans. The mixed symptoms caused byBecause of its potential application the report was widelymycoplasmas are not the same as caused by bacteria and viruses.publicized in newspapers and magazines. For example theThe different responses are attributed to mycoplasma’s differentAssociated Press reported “Hope Held out for Cure of Rheumaticphysical, chemical and host related properties.Ills, Washington research team announces New Concept on CausesWhat is Mycoplasma Arthritis?and Cures”. At that time except for the veterinarians, very fewThe common symptom in most Rheumatic Diseases isdoctors knew what a PPLO or mycoplasma was, let alone its role ininflammation as anatomically described by synovitis, bursitis,human arthritis.carditis, neuritis, or nephritis, etc. Many different causes ofAlthough Koch and Pasteur had brought bacteria to theinflammation have been suggested and pursued. Upon furtherforefront of medicine it was the microbiologists Nocard and Rouxstudy and review, researchers may find that a common microbe,at the Pasteur Institute in 1898 that first isolated a filtrable bacteriasuch as Mycoplasmas can cause various host responses orfrom pneumonic and arthritic cattle. For the next 65 years new1

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.symptoms in similar tissues. Because of the different portals ofentry differences between individuals may be observed. Because ofthe associative factors; age, gender, genetics, and environment amicroorganism could cause arthritis in one patient, systemic lupuserythematosus (SLE) in another, or neither disorder. The sameagent and mechanism that causes inflammation of the synovialtissues in a male patient may cause nephritis or both in a female. Asin Juvenile Rheumatoid Arthritis, the juvenile host could responddifferently to mycoplasmas than a mature host. Mycoplasmascould infect the fetus and lay dormant until challenged and latercould cause different symptoms.Of all the targeted organs and tissues, the blood supply is trulyour lifeline and first line of defense that controls the severity andspread of viable agents. The blood supply may determine theprimary site of disease activity, such as the kidney or liver, orsynovium, and whether it is localized or widespread. Some targetedtissues such as the synovium, eyes, and kidneys are morevulnerable to the inflammatory disorders because they are morevascularized and present symmetrical symptoms. The compositionand restriction of the blood supply by inflamed and swollen tissuesbecomes life-threatening if they control the blood pressure andvolume. The potential role of mycoplasmas in the formation ofarterial plaques and the subsequent host responses must also beconsidered. The availability of receptive tissues with attachmentsites for a particular mycoplasma strain are essential for the hostresponses.We should ask: How do Mycoplasmas cause Inflammation?In 1983 the USA National Arthritis Advisory Board reportedthat“Because we know that mycoplasmas can cause arthritis inmany animals and because we know that they do cause acute andchronic disease in humans (in lungs, and genitourinary tract) wemust take seriously the possibility Mycoplasmas cause arthritis inhumans.” As in arthritic animal models (pigs, rats, mice) thedetection of viable mycoplasmas in arthritic patients “now you seethem, now you don’t” has made “Hocus-Pocus” skeptics of manydoctors when arthritis symptoms and mycoplasmas seem to hideand come and go with the weather. As in animal modelsmycoplasmas are difficult to isolate or recover even though theirarthritis and serologic response continues. Unlike viruses,mycoplasmas can be isolated and cultured in liquid and on solidcell free media and unlike bacteria mycoplasmas require preformedmacro molecules from digested tissues. The “fried egg” colonymorphology of mycoplasmas on agar plates is characteristic but notalways typical and depends on both the culture and agar media asshown in this slide, from tissue cell culture. (See Figure 1A & 1B)By using a hot water fixation technique, the stickymycoplasma colonies can be fixed on a slide for microscopicexamination free of agar and bacteria. The technique consists ofplacing an inverted agar block on a glass slide that is immersed uprightin a beaker of hot (80 0 C) water dissolving the agar leavingattached mycoplasma colonies intact as shown in Figures 2A & 2B.The variable colony sizes are dependent upon the aggregatedminimal reproductive units in the inoculum as well as the agarmedia. M. pneumoniae adhere to glass culture flasks requiringscraping to recover. The M. pneumoniae agar colonies, shown inFigure 1, are both large and small. Shadowed with oblique lightgives an added dimension. Investigators identified this newmycoplasma strain by serologically testing the hot water fixedcolonies on a slide using fluorescent labeled antisera developed inour lab ( J. Bact. ‘63). Greater details of stained hot water fixedcolonies show their particle composition when stained with Giemsaor for lipids and nucleic acids.See Figure 2A: A dark-field photo of a mycoplasma colonyshowing the minimal reproductive particles.See Figure 2B: Shows the adhesiveness and pliability ofmycoplasmas from a colony streaked on a slide producing filamentlike structures.Most people know what arthritis is but only a few know aboutmycoplasmas, a unique microbial cause of arthritis in animals andin humans. Besides being the smallest free-living microorganisms,(between 50-100 nm), mycoplasmas without a cell wall are highlypleomorphic and likened to a viral size jelly-fish. Unlike the largerand more parasitic bacteria and smaller viruses, mycoplasmas areboth saprophytic and parasitic opportunists living off the cellulardebris as in tissue cell cultures or complex cell-free media.See Figures 3A & 3B: Mycoplasmas such as shown in theelectron photomicrographs are rarely captured in a natural formbecause of their pliable and sticky lipid membranes when using theold formalin protein fixative. The Mycoplasmas were grown inserum enriched cell-free broth, diluted in buffer, centrifuged on toEM grids and cautiously fixed with dilute osmium tetroxide andshadowed to capture their true structure and reproductive cycle asshown by the hexadic budding of six minimal reproductive units.Note chain of six budding rings.Electron photomicrographs of shadowed mycoplasmaspecimens show donut (ring) shaped mycoplasmas at low and highmagnification.Rings or cherios, dented open from sedimentation, result incollapsed cell membrane. Mycoplasma cells appear flat when airdried. Culture of M.gallisepticum mid-stage growth, shows ringswith 6 buds.Similar morphology and division cycles were observed in1935 by Dr. Nobel.See Figures 4A & 4B: Shows the variable sizes and shapes ingrowth cycle, like collapsed balloons, and a single mycoplasmaring or donut,The three basic stages of growth with hexadic (6) budding ofthe minimum reproductive units, and with chains of budding rings.The most significant observation of mycoplasma morphologyand growth cycle is their hexadic budding into six viral sizereproductive units that was reported in 1965, J. Bact. and again in1985. This division may vary in other strains.The basis for this hexadic budding was recently indicated byGerman investigators 1997, who were sequencing the genomes oftwo mycoplasma strains, pneumoniae & genitalium. They foundthat the genomes of both mycoplasmas could be subdivided into sixequal segments. The other genes were well conserved within eachof the individual segments but the arrangement of the six segmentsdiffered. This hexadic growth cycle now with a genomical basisshould be pursued in the host’s cellular pathology and observed inthe host’s tissues.See Figure 5 A: High magnification showing membranesurface with adhesive attachment and infrastructure.See Figure 5 B: Mycoplasma’s sensitivity to low osmolarityare morphology and viability dependent, showing coccoidal inisotonic saline, and filamentous in water.The fixation of the lipoprotein membrane with OsmiumTetroxide is in marked contrast to the segmented filamentous formsobserved in the formalin fixed specimens. Some mycoplasmaspecimens seem to show their infrastructure and composition. Theneed for a closer examination of all mycoplasma strains comparingboth fixation techniques is apparent as their morphology andgrowth cycles may be characteristically different. For example: M.gallisepticum has a greater growth rate than M. hominis.Mycoplasma’s minimal reproductive unit size as determined2

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.by ultrafiltration is less than 100 mu and dependent upon the strainand age of culture.The filtration pressure could force larger pliable particlescomposition.Each strain has its own antigenic pattern providing serologicspecificity with polyvalent antiserum.through the millipore membranes. With an estimated mass of 1.2 x See Figure 7A: The antisera labeling of the SDS10 -10 ug. the smallest reproductive unit (bud) the genetic codingDNA would be approximately 4 x 10 6 molecular weight andsuggests a limited synthetic capability. (99% larger than 300)CompositionIn 1962 we were awarded a grant from the NIH to investigatemycoplasma’s chemical composition. This was the beginning of anendless problem. In addition to the wide ranging cholesterolcomposition we found the protein, nucleic acid, and lipidelectrophoresis of mycoplasma antigens show M.salivariumantigen (11) preparation contaminated with common mediaantigens found in other strains. fermentans (10) & orale (12).See Figure 7B: The laser scan of the SDS electrophoresis blotof M.salivarium, top: shows silver stained organic composition,middle: antigens reactive with antisera and bottom controlshowing non-specific media antigens observed in other strains.Like their human and animal hosts mycoplasmas mimic theircomposition varied with changes in the tissue broth digest and culture media, “they are what they eat.” For exampleserum enrichment. Little known and rarely reported, the basicpeptides from tryptic tissue digests are essential for mycoplasmagrowth and apparently can reflect its tissue source. In addition thebasic proteins such as IgG from serum enrichment was found to beincorporated by mycoplasma during In-Vitro growth. Thus itseems probable that mycoplasmas in the human host couldincorporate and alter specific basic proteins in any number oftissues and form autoantigens but also act as their adjuvant carrier.A basic protein such as IgG becomes altered when attached to themycoplasma carrier thus rendering it autoantigenic to the host.Native IgG or a basic protein is nonreactive to the human hostunless attached to adjuvant. The autoimmune rheumatoid factor,anti-antibody, is only reactive to the IgG that is altered byattachment to a particle such as mycoplasmas, Rbc, or laytex. Thespecificity of basic peptides or proteins may depend on themycoplasma strains with affinity to certain tissues indicated bytheir differences in other metabolic activity.Digested broth extracts prepared from different tissues werefound to equally support mycoplasma growth. The resultingmycoplasmas were found to have different composition andantigenic properties. Although the techniques available todaycould provide further details I believe the results would remain thesame.A marked difference was found in the phosphorus and nucleicacid composition of mycoplasmas cultured in a peptic digested beefbroth (PD) and a tryptose digested casein-soy broth (TS). .A marked decrease in the DNA composition was found inmycoplasmas cultured in media containing penicillin. Noninhibitingantibiotics penicillin and ampicillin appreciablydecreased mycoplasma’s antigenicity when incorporated in theirculture media. Other factors such as pH, and culture age couldcause composition changes.Mycoplasma’s antibiotic sensitivities are also altered. Inanother experiment using different media and strains thedifferences in media and mycoplasma composition were lessmarked.Complement Fixation (MCF) titers in immunized guinea pigsshowed mycoplasmas cultured in digested thymus broth morereactive and lung tissue specific.Variation in mycoplasmaantigenicty was found in different culture media plus or minusantibiotics.See Figure 6A (Table 6A): The mycoplasma complementfixation titers in sera from patients in a respiratory disease clinicwas more reactive with mycoplasmas cultured in lung digest brothagain indicating autoimmune reactivity and mixed infections. Lowantigenicity in standard BH media. 117 positive for M. hominis.See Figure 6B: Disc-gel electrophoresis shows differences inprotein composition of mycoplasmas cultured in different tissuedigest broths from human sources.Casein Soy and Intestine digest broths show different proteinmycoplasma’s cholesterol levels are elevated when grown in a highcholesterol media, that is essential for some strains as part of theirlipoprotein membrane. Complicating research further, mycoplasmasmimic their media and change their properties depending ontheir localization in tissues: brain, lung, kidney, heart, synovium,In-Vitro. How much surplus food can and do mycoplasmas ingest?Mycoplasmas seem to grow equally in diluted (10x) andconcentrated broth and more apt to colonize and infiltrate tissueswith the highest degenerative rate. Unlike most inflammatoryagents mycoplasma’s unique “Sleepy Virus” properties make thema prime candidate as persistent viable antigens and a causative rolein different chronic autoimmune diseases; such as rheumatoidarthritis, lupus, diabetes, MS, etc.What is your blood cholesterol level?Mycoplasmas, like humans, not only require cholesterol forgrowth but can incorporate excessive concentration growing in ahigh cholesterol media -- such as a high blood level. What we don’tknow but suspect is mycoplasma’s role in arterial plaque formationand the adverse effects of their elevated cholesterol levels.Besides their specific requirement for basic peptides fromtissue protein digests, mycoplasmas also require the nucleic acidprecursors, purines and pyrimidines. These may be provided by theRNA and DNA nucleotides that have been degraded by themycoplasma’s nucleases. Although mycoplasmas generallysynthesize their own membrane phospholipids and lipoproteinsfrom the exogenously provided fatty acids, some strainsincorporate preformed host phospholipids into their membranes asanother autoantigen. Being deficient in their ability to regulate themembrane fluidity by preferential fatty acid biosynthesis, themycoplasmas overcome this difficulty by incorporating largequantities of exogenous cholesterol into their membrane.Cell Membrane composition.Proteins constitute over two-thirds of the mycoplasmamembrane with the rest being lipids. The membrane lipoproteinshave attracted much attention since their abundance is remarkablein contrast to the limited number in eubacteria. Membranelipoproteins are among the most dominant antigens in mollicutesand known to undergo antigenic variations.Based on characteristic lipoprotein specific features, Germaninvestigators counted 46 lipoprotein genes in the M. Pneumoniaegenome, while 21 putative lipoprotein genes were found in the M.genitalium genome. In this regard a lipoprotein extract fromspinach has been used as the antigen to serologically test for M.pneumoniae antibodies indicating a broad range of lipoproteinantigens.Virtually all mycoplasma lipids are located in the cellmembrane and as in other biological membranes, consist ofphospholipids, glycolipids, and neutral lipids that constitute amajor portion of the hydrophobic core. Mycoplasmas are incapableof fatty acid synthesis and depend on the host or culture medium for3

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.their supply, including cholesterol. The dependence of autoimmune antigens, especially when the haptenic basic peptidesMycoplasmas on the exogenous supply of fattty acids andcholesterol has been one of their greatest advantages as models formembrane investigations. Thus the basis for the Osmium Tetroxidelipid fixation.Due to the lack of a cell wall, the mycoplasma cell membraneis exposed to the external environment. This can facilitate directcontact of the mycoplasma membrane with its eukaryotic host,which could lead to fusion of the two membranes enabling thetransfer and exchange of membrane components.Thus the fusion or attachment of host membrane componentsis a potential source of antoantigens.Energy MetabolismPrimarily because of their small size the mycoplasmas possessthe minimum machinery for protein synthesis and growth.Consequently mycoplasma cell replication is much slower than inbacteria. Most striking is the lack of many energy-yielding systems.required for growth induce tissue specific responses. Some strainsof mycoplasmas are known to cross-react with different tissuesubstances as an autoimmune response.10) Localized and persisting mycoplasma antibodies arefound in the synovial fluid of RA patients. The titers of circulatingmycoplasma antibodies frequently are inversely correlated withthe titer of rheumatoid factor (anti-antibody) .11) Mycoplasmas participate in several cell -mediatedresponses particularly the immunoinflammatory reactions of thedelayed-type skin test.12) The immunosuppressive effect of mycoplasmas on thereactivity of peripheral T lymphocytes is comparable to thesuppressor activity associated with autoimmune responses.13) Without a cell wall mycoplasmas are inhibited by avariety of substances used to treat RA, i.e. antimalarials, gold salts,bee venom, and tetracyclines.No tricarboxylic acid cycle, and no quinone and no cytochromehave been found in any of the mycoplasmas. The irony here is thatafter spending 4 years in graduate school investigating therespiratory cytochrome chain, I end-up spending 35 yearsinvestigating microbial pathogens that are cytochrome deficient.The electron transport system in mycoplasmas is flavin terminated.Thus high energy ATP is produced in mycoplasmas by substratelevel phosphorylation, a less efficient mechanism than oxidativephosphorylation. Most strains are facultative anaerobes andantioxidants could impede their growth.Some investigators have added high energy ATP to the culturemedia.Summary of Mycoplasma’s PropertiesThe unique properties of mycoplasmas supports their role asetiologic agents in Rheumatoid and Autoimmune Diseases.1) Mycoplasmas are recognized as arthritogenic and systemicagents in animals and humans, with strain and species specificity.2) Colonization of both nasopharyngeal and genitourinarytracts by mycoplasmas is common, infecting four times as manywomen as men, a predilection in autoimmune diseases thatpredominantly affect females.3) Several strains isolated from the same and different animalspecies have marked physical, chemical and pathogenic variations.4) The pleomorphic characteristic of mycoplasmas, the resultof the lack of a cell wall, their unusual growth cycles and growthrequirements contribute to the problem of isolating and identifyingmycoplasmas in host tissues.5) The low cytotoxicity of some strains of mycoplasma thathave gone undetected in tissue cell cultures would support thefinding of a low-titer antibody response and persistence of themycoplasmas in symptom free hosts.6) Finding that the greatest incidence of mycoplasmalantibodies in humans associated with other infections wouldindicate that some mycoplasma strains are commensal or secondaryinvaders.7) Mycoplasmas are difficult to eliminate from their infectedhost with antibiotics and vaccines, a factor that indicatesmycoplasmas persist fixed in or on tissues. Such persistence couldaccount for the infrequency with which the organisms are isolatedfrom host tissues.8.) The rise in antibody titer to specific mycoplasmas and adelayed-type inflammatory response following trauma or therapyindicates the release of a persistent antigen, such as a Herxheimerreaction in a sensitized host.9) The characteristic of mycoplasmas, who’s compositionmimics the culture media, could contribute to the production ofLecture II: Lecture on Recent Therapies of Human MycoplasmaArthritis at the San Marcos University College ofVeterinarian Medicine in Lima, Peru. (Summary)What is mycoplasma arthritis and how do we effectively andsafely treat the host?Mycoplasma Arthritis -- as defined by the ArthritisFoundation Primer on Rheumatic Disease, 10th Ed, 1993,“Mycoplasma pneumoniae infection is often associated with nonspecificarthralgias and myalgias. In some patients a migratorypolyarthritis of medium size joints such as the shoulders, elbows,knees, and ankles may occur three to eight days after onset ofinitial illness and may last two months. In a few patients symptomsmay last up to one year.” Mycoplasma arthritis is now listed as oneof the 171 types of arthritis.In the Beginning:Over 4 thousand years ago, in Egypt, the high priests treatedArthritic Consumption in the Pharoes with beer and bread madefrom moldy grain. Little did they know that the streptomyces moldon the grain and bread was a source of today’s most effectivetetracycline antibiotics.In today’s modern civilization of purified bottled waterEgyptian women seem to have the highest incidence of Lupus, asevere autoimmune disorder and rheumatic disease also known asImmune Complex and Collagen Vascular disorders. Even thoughthe causative agent may be the same, the symptoms of rheumatoidarthritis and other rheumatic diseases are not necessarily the samearound the world. Most of our chronic diseases continue to persistwhile the causes remain unknown due to our genotypicdifferences. Having elevated medicine from willow bark toaspirins and from cinchona bark to quinine alkaloids, the failure oftoday’s health care, in desperation, seems to be returning back tothe beginning with herbs and food supplements or back to nature.Many new symptomatic therapies have failed with time. Just asmany of the infectious diseases, Tb, leprosy, and small pox werereduced in the world until something hidden like AIDS rises to thesurface. We should also consider whether chlorinating andfluorinating our drinking water, while limiting some diseases mayopen the door for other diseases, like cancer, neurologic, and heartdiseases. (See “Flouridation: Governmentally Approved Poison,”http://www.arthritistrust.org. Ed.) By eliminating the infectiouscause of rheumatoid diseases we could also reduce the incidence ofother diseases and premature deaths. Therapies that reduce oreliminate symptoms by inhibiting one oxidizing enzyme (Cox2)could also open the doors to the inhibition of other essentialoxidases causing an adverse break in metabolic chains. Tissues4

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.smothered with excessive antioxidants may not survive and the antimicrobial. With intermittent dosing there is less chance ofover use of immunosuppressives can open doors for more severe causing resistant strains and toxicity. There is the precedent ofinfections like a double edge sword. Pulse therapy will allow the other therapies, gold and methotrexate where weekly pulse dosestissues to catch their breath, especially at higher elevations. were found to be less toxic and thus more tolerable and effectiveWhereas the increased arthritis swelling and pain at that low over time. (See “The Roger Wyburn-Mason Treatment forpressure altitude could be attributed to diving down in the ocean Rheumatoid Disease,” “Foreward,” http://www.arthritistrust.org.where the increase pressure could reduce the volume or swelling Ed.) Even the one day pause allows recovery of the host tissuesand pain.from antioxidants, immunosuppressants, and protein synthesisIn view of the thousands of therapies proposed for arthritis the inhibitor effects. Treatment is more effective chipping away overfocus should be primarily on those aimed at eliminating and long periods with the eventual inhibition of the viable and lesspreventing an infectious mycoplasma cause and not just symptoms. virulent mycoplasmas rather than overwhelm the tissues with drugMuch like the Tb mycobacterium, mycoplasmas are difficult to toxicity. In most chronic diseases a drugs toxic actions on the hostisolate and eliminate even with multiple antibiotics.tissues is more critical than its effect on a benign causativeA Mechanistic Treatment of Mycoplasma Arthritis microbe.A mechanistic approach to the treatment of the infectious Chelated tetracyclines fixed in bone and other low metabolicrheumatoid diseases can be more effective and less costly by tissues like cartilage provide a pool, replenished by intermittentcontrolling and preventing the disease and not just the symptoms. dosage. Protein inhibitors’ excessive long term doses can damageIn addition to sustaining good health basically there are three organ functions and negate any benefits. Having to discontinuetargets:gold salts therapy after a few years is a good example. Antibiotic1) to SEARCH for and eliminate a persistent irritating effectiveness is dependent on the tissue infected. For examplemicrobial cause(s).mixed I.V. and oral dosages may be required to clear the fatty2) to identify and BLOCK the antigen-antibody immune synovium tissue unlike the genital tract. In-Vitro studies foundcomplex formation, andthat mycoplasma’s antigenicity and antibiotisensitivity were3) to control and ELIMINATE enzyme induced inflammation variable when cultured in different tissue broths indicating thewith tissue destruction.need for variable antibiotics and dosages. Bacteriostatic bloodThe elimination of the microbial root cause should be the levels are not always critical when evaluating a multi action drugprimary target. The less pathogenic or nonvirulent microbes should such as the tetracyclines.be less immunologically reactive and require less antibiotic Even with specific genetic coding the complex pathogenicitytreatment.of mycoplasmas become a set of conditions that regulate the hostparasiteor saprophytic interaction. A pathogenic substance suchThree Prong Prescription >>> adjusted for each patient1. Antibiotics: such as minocyclines, in low pulsed doses as mycoplasmas that cause a delayed or persistent abnormal hostshould be directed at eliminating and inhibiting the atypical response in one set of conditions may be nonpathogenic in anothermicrobial cause. In addition the three-prong tetracyclines can also set. Some strains of mycoplasmas and ureaplasmas are speciesact as Antioxidants, Immunosuppressants, protein synthesis and specific for humans and animals and are specific pathogens formetaloenzyme inhibitors.respiratory and genitourinary tract tissues. However in tissue cell2. Immunosuppressants: such as prednisone, and many other cultures the mycoplasmas were found to coexist as cell boundchemicals that block the immune-complex formation that activates noncytopathogenic saprophytes and remain untouched byComplement proteolysis and further promotes tissue destructive antibiotic or antisera treatment. After 50 years of recognitioninflammation.mycoplasma’s continued persistence in tissue cell cultures3, Antiinflammatory antioxidants: such as the non-steroidal indicate their resistance to therapeutic control in human tissues andantiinflammatory drugs (NSAIDS and Cox-2) to eliminate and the need for a special approach.prevent the tissue destructive inflammation.Another form of mycoplasma pathogenicity is their specificIn addition the essential role of several major contributing interaction with the host’s basic proteins such as IgG with theirfactors affecting the occurrence and severity of the chronic arthritis selective adhesion and conformation of specific cell membraneand immunologic disorders must also be considered in selecting autoantigens. The cell bound immune complexes (IC) ofand benefiting from the most effective treatment. These include antigen+antibody have been cited as pathogens for rheumaticvariable factors such as; physical and mental Health, Exercise, diseases. An example is finding mycoplasma antigens in the ICNutrition, Allergies, Exposure, physical and mental Stress. The extracted from rheumatoid synovial fluids.decisive fixed factors include: Age, Gender, and Genetic The mycoplasma may have been transmitted in-utero and laysucceptability, all of which can help hinder or predispose the dormant until the host cells become receptive. The disease or thetherapeutic success. These three therapeutic components are most host response also depends on the pathogen’s portal of entry. Theeffective when used collectively. The probes are only as effective foreign antigens could elicit pathogenic host responses such asas the recipient’s good health that can provide a unified basis for leukocyte activation resulting in antibody production andsafe and effective therapy individually customized for each patient. inflammation with tissue destruction. Also the attachment ofEven the tetracycline’s lipophilic, nucleophilic and chelating mycoplasmas to blood cells, lymph vessels, and nerves could beproperties that have positive multi-directional actions, must focus responsible for plaque formation and impaired circulation as withsimultaneously on all and not just one action, as antibiotics can act the resulting agalactia restriction of milk in goats and thedifferently in each patient. (Also see “Allergies and Biodetoxification restriction of capillary fluids in plant and tree wilting diseases byfor the Arthritic,” “Stress,” “Proper Nutrition for Rheumatoid the related citroplasma strains.Arthritis,” http://www.arthritistrust.org. Ed.)Mycoplasmas present a variable target as both extracellularTreatment: for Chronic Mycoplasma Arthritis and intracellular agents. With low cytopathogenicity, mycoplasmaIntermittent dosing is important especially with drugs such as can remain attached to and lay dormant in tissue cells for monthstetracylines that are multiple acting, like aspirin, and not just5

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.without killing the cells as long as the cells continue to shed antibiotic treatment is given at the early onset of pneumonia as innutrients. For years mycoplasmas (PPLO) were primarily rheumatoid arthritis. Even after treatment M. pneumoniae is shedconsidered nonpathogenic in humans even though veterinarians indicating a hidden source of infection. Being hemolytic andknew of their severe pathology in animals. Finding a closely related reactive with specific red blood cell membrane antigens M.gorilla model with severe spontaneous rheumatoid arthritis and pneumoniae elicits cold agglutinin antisera and autoimmunescheduled for euthanasia provided the ideal test subject. The model reactions.was an under weight five year male infected and serologically Based on positive serological blood tests females were foundpositive for a human Mycoplasma salivarium strain. He responded to be infected with mycoplasmas 4 times more frequently thanslowly as expected, initially with a Herxheimer reaction to the males, a prerequisite to mycoplasma’s role in the female dominant1,000 mg IV doxycycline bi-monthly, resulting in less frequent autoimmune rheumatoid disorders. Persons with greater exposureand diminished flares. Over 4 years of treatment his weight tripled especially the females were found infected more. Using severaland he became a beautiful silver back gorilla specimen. Providing mycoplasma strains our lab found clinic patients up to 80%culture and serologic tests for mycoplasmas in captive gorillas in positive.zoos around the world we learned of other arthritic gorillas who Even though the viable mycoplasmas are frequentlyalso responded to the I.V. tetracycline treatment.transmitted among families, arthritis in humans usually is notArthritogenic mycoplasma have been investigated and contagious. However, because of possible family contact andfrequently reported in domestic animals such as; bovine, caprine, mycoplasma transmission, antibiotic treatment of the familyswine, rodents, avian, etc. Many others with species specific should be considered especially if they are both infected. As inmycoplasma strains, including; canine, feline, equine, elephants, other chronic diseases the genetic or family susceptibility as well asetc. have been isolated but seldom associated with their arthritis or poor health and stress all contribute to the host’s symptoms andantibiotic treatment. As in humans the experimental animal models response to treatment. The apparent increase in femaleshow the difficulty in isolating or treating mycoplasma arthritis susceptibility to mycoplasma infection may also indicate a need forespecially in the late stages.higher antibiotic dosage in treating mycoplasma arthritis inAgain I.V. antibiotic treatment has been found more effective females. Unfortunately when the mycoplasmas acquire componentsand properties from the host’s tissues they become less of anthan oral but more difficult and costly to administer.Mycoplasmas, a.k.a. PPLO, were first isolated from a human irritant and as an autoantigen result in abnormally weak immuneovarian abscess in 1932. Although strains are frequently isolated responses that are more difficult to measure.from the respiratory and urogenital tracts their isolation from Four years ago several clinical trials confirmed that thesynovial fluid and tissues has remained infrequent. Mycoplasmas antibiotic minocycline was a safe and effective treatment ofand Ureaplasmas, a related species, have been isolated from and or rheumatoid arthritis. These results supported the suggestion thedetected in the broad spectrum of systemic tissues. Even though late Thomas McPherson Brown, M.D. made in 1949 that antibioticfrequently associated with their host, identifying their role in therapy may be beneficial for some people with RA that was causedarthritis or other rheumatic diseases still remains. Using today’s by an infection with a microorganism called a mycoplasma in thetechniques the frequency of mycoplasma isolation and association joints, and that oral and/or intravenous antibiotics should be used inhas been greatly enhanced..the disease’s treatment.Ureaplasmas have been found associated with natural and The book Why Arthritis? By Harold W.Clark answers theexperimental inflammatory disease in both animals and humans. In questions, “Why has it taken over 40 years to test and demonstratehumans NG urethritis and arthritis are the primary symptoms antibiotic’s effectiveness?” and “Why withhold a safe and effectiveassociated with some strains of ureaplasmas. Because of their treatment when nothing safer and more effective is available?”similarities mycoplasmas could be considered a mixed infection Although the 48 week clinical trial of oral minicycline on twoidentified by a mixed culture with tiny ureaplasma colonies. In hundred rheumatoid arthritis patients was found to be safe andaddition to arthritis, respiratory, neurologic, and urogenital effective there apparently was no attempt to measure thediseases, ureaplasmas and mycoplasmas have also been found occurrence of any infectious agents before or after treatment.associated with spontaneous abortion, infertility and low Several clinics world wide have shown minocycline treatment bothbirthweight in both animals and humans that have frequently been safe and effective in the treatment of rheumatoid arthritis. In viewcontrolled with tetracycline therapy. Ureaplasmas have also been of this it is discouraging to see the Arthritis Foundation listfound in the fetal brain and spinal fluid, and from children of all antibacterial treatment under “Questionable Therapies” togetherages as well as adults, especially those sexually active and child with many unproven remedies in their current 1999 Primer onbearing age. The appearance of rheumatoid arthritis following the Rheumatic Diseases. Minocycline is listed as a disease modifyingdelivery of the second child could indicate a delayed antirheumatic drug (DMARD) but not recomended for treatment.hypersensitivity response to previous contact and following an A five year comparison of the effectiveness of the approved goldimmunosuppressive period. Thus in some cases mycoplasma could salts therapy with the much less toxic antibiotic therapy ispersist as a slowly developing irritant finally surfacing years later in illustrated in Figures 8 & 9.chronic inflammation symptoms of arthritis and other autoimmuneWhen Mycoplasma Arthritis is listed as one of the 171diseases. Unless the host’s immune defense system is mature and types of arthritis, antibiotic treatment should be essential and nothealthy and brings the mycoplasma infection under control, questionable. Obviously any infection associated with arthritis, inadditional help with antimycoplasma therapy should be prescribed view of its unknown cause, should be treated with antibioticsfor both their control and prevention at the earliest possible time. especially if it has been proven safe and effective. Failure to test forThe clinical spectrum of M. pneumoniae diseases is far from and identify Mycoplasmas or other infectious agents does notcomplete. Originally thought to be a unique respiratory virus exclude their causal role in rheumatoid or autoimmune diseases.specific for humans and hamsters it has been associated with Also the safe and effective response to antibiotic therapy does notmusculo skeletal arthritis as well as neurologic G-B syndrome, prove an infectious cause. Unlike mycoplasma arthritis, syphilitic,hematologic and respiratory diseases. To be most effective tuberculosis, or gonococcal arthritis are highly contagious systemic6

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.diseases where arthritis is only one of the many symptoms that palliation of symptoms.require special therapeutic control.Why test for Mycoplasmas?From the book: Why Arthritis? Searching for the Cause andSome atypical viral-like infectious agent has long been the Cure of Rheumatoid Disease by Harold W. Clark, Ph.D.suspected of causing chronic rheumatic diseases but rarely tested http//www.hitter.net/users/hwcmriwith antimycoplasma treatment. Testing and finding mycoplasma,although important, is not easy and requires a research lab withspecial skills to do acceptable tests. Testing for mycoplasmaantibodies and antigens can be done three ways: by culture,Polymerase Chain Reaction, or serologic. Patients can havemultiple infections, many of which may be involved in therheumatic disease process. The multiple strains of mycoplasmasincluding the ureaplasmas can respond to different antibiotics. Ifboth strains are present 2 antibiotics should be given to eliminateboth strains. As mentioned tetracycline and other antibiotics havemultiple actions that can give relief of symptoms independent oftheir biostatic sensitivity levels. A positive test for mycoplasmas orother microbial infection requires the appropriate antibiotictreatment for the primary diagnosis and arthritis as the secondarydiagnosis.Many of the rheumatoid diseases are now recognized asimmune complex (IC) disorders, the result of binding microbialantigens with their antibodies.The IC formation, fixed in tissues, activates the complementcascade of proteolytic enzymes causing synovitis, vasculitis,dermatitis and glomerulonephritis. What is still not recognized andaccepted is the multi action of tetracycline antibiotics asimmunosuppressants in blocking the IC formation. Except for theirtoxicity other beneficial medications, (NSAIDS,cortisone) couldhave a similar but limited effects.Non-RespondersAlthough many patients with various rheumatoid diseaseshave responded well to long-term antibiotic therapy, there are somewho do not respond or stop. There can be several reasons for apatient’s failure to fully respond. Some of these include: Somegeneric brands do not work at the same dose and frequency as thedrug tested in original trials. Other pre-existing conditions andmedications may hinder the response to antibiotic treatment. Thepatient’s immune system may be deficient or stressed caused by aconcurrent infection or condition limiting the attack onmycoplasmas. The patient is self prescribing over-the-countermedications that are contra indicated for use with tetracyclines,such as vitamins, minerals and antacids. Also hormonal ormetabolic imbalance could interfere with antibiotic action. Somestrains of mycoplasmas may be found to be resistant to someantibiotics requiring a change to one more sensitive. Poor gutabsorption may limit medications effectiveness. The inflammationmay cause a barrier to drugs effectiveness requiring a constantsupply of antiinflammatory (NSAIDs) medication. After years oflong term antibiotic therapy a patient could become tolerant of theantibiotic requiring a change. The possibility of a recurrentexposure to mycoplasmas must be eliminated. The dosage andantibiotic must be adjusted, higher or lower for each patient overtime to achieve maximum benefits. Intravenous and oral treatmentmay be required. Long term therapy response is usually slow andgradual with rate and extent of improvement depending on theduration and severity of disease. The patient’s response tomedication is very individualized, male and female, young and old,with treatment adjusted frequently to accommodate their needs.In Conclusion:I will leave it up to the patients and their doctors to decidewhether treatment should include searching for and treating aninfectious immunological cause and not just the continued7

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Mycoplasma ColoniesSlide1BFigure 1B: On Agar Plate, 100X.The “fried egg” colony morphology of mycoplasmas onagar plates is characteristic but not always typical and depends on both the culture andagar media as shown in this slide, from tissue cell culture8

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 2A: Mycoplasma Colony, Fixed on Glass Slide, 400X dark field.A dark-field photo of a mycoplasma colony showing the minimal reproductive particles.9

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.2B. Dark-field photomicrographs of hot water-fixed Mycoplasma hominis, type 1, colonies,X 450. Variable large body and particulate chainlike filaments disrupted from colony.Shows the adhesiveness and pliability of mycoplasmas from a colony streakedon a slide producing filament like structures.10

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 3A: Electromicroscopic view of broth culture of Mycoplasma hominis, Type II,isolated from patient with Rheumatoid Arthritis X 20,000 magnification.Note bizarre, non-specific, poorly identified appearance of microbial elements.11

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 3B: Published: Journal of Bacteriology 1965, 90, 1373-1386;Mycoplasmas -- hexadic (6) budding12

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 4A: Mycoplasma: 100,000 magnification13

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 4B: Mycoplasma Culture: Electron Microscope14

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 5A: Five Mycoplasmas: 120,000 X magnification by electronmicroscope. Highmagnification showing membrane surface with adhesive attachment and infrastructure.15

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 5B: Mycoplasma in water and salt. Mycoplasma’s sensitivity to low osmolarity aremorphology and viability dependent, showing coccoidal in isotonic saline,and filamentous in water.16

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 6A (Table 6A): Influence of Culture Media on Mycoplasma Complement Fixation(CF) Activity in the Sera from 48 Pulmonary Disease Clinic Patients*Mycoplasma antigensM. M. M. M.M. orale (Patt) M. salivarium (PG-20) pneumoniae hominis arthritidis fermentonsMedia B P-lung H-lung B P-lung H-lung B B B BCFpositive 2 6 13 11 10 0 2 17 6 0Symbols: B = Bovine heart; P = pig; H = human. 29 = (+), 1:8 or greater; 19 = (-).* Harold W. Clark,Jack S. Bailey, and Thomas McPherson Brown, “Medium-dependent Properties of Mycoplasmas,” Diagn Microbiol Infect Dis,3:283-294, 1985.The mycoplasma complement fixation titers in sera from patients in a respiratory diseaseclinic was more reactive with mycoplasmas cultured in lung digest broth again indicatingautoimmune reactivity and mixed infections. Low antigenicity standard BH media.117 positive for M. hominis.17

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.BDisc-gel electrophoresis shows differences in proteincomposition of mycoplasmas culturedin different tissue digest broths from human sources.18

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 7A: The antisera labeling of the SDS electrophoresis of mycoplasma antigens showM.salivarium antigen (11) preparation contaminated with common media antigens found inother strains. fermentans (10) & orale (12).19

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.Figure 7B: The laser scan of the SDS electrophoresis blot of M.salivarium, top: showssilver stained organic composition, middle: antigens reactive with antisera and bottomcontrol showing non-specific media antigens observed in other strains.20

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.21

Medical data is for informational purposes only. You should always consult your family physician, or one of our referral physicians prior to treatment.22