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ASSISTEB REPRODUCTIVE TECHNOLOGY-MALEJSA PRESENTATIONSTHE ROLES OF SOX GENES IN MARSUPIAL SEX DETERMINATIONTHE ENTRY OF IMMUNOGLOBULIN INTO THE MALE REPRODUCTIVETRACTH. Ecroyd\ K.W. Beagle/, N. Newcombe' and R. C. Jones'.lDepartment ofBiological Sciences, University ofNewcastle, NSW 2308 and2Discipline ofPathology, Faculty ofMedicine and Health Sciences, The University ofNewcastle, NSW 2308INTRODUCTIONThe male reproductive tract is often considered animmunologically privileged site due to the presence ofboth anatomical (1,2) and immunological barriers thatact to shield reproductive antigens in the duct lumen.Knowledge of the local immunity of the malereproductive system and its regulation (both locallyand systemically), is fundamental to studies in anumber of fields. This includes the development ofvaccines against male STD's and maleimmunocontraceptive vaccines. The success of suchvaccines is dependent upon the induced antibodiesaccessing the target antigen in sufficient titres as to beeffective. The aims of the present study were toestablish the site(s) of entry of specific 19A and IgGantibody into the duct lumen ofthe male reproductivetract and to determine immunisation regimes thatmaximise their delivery.MATERIALS AND METHODSMale Balb/c mice were immunised with tetanustoxoid (IT) by subcutaneous, oral, intranasal (IN),rectal or transdermal routes. Luminal reproductivefluids were collected from the rete testis (RT), caudaepididymidis (CE), seminal vesicles and ventralprostate (P) gland and specific anti-tetanus IgG andIgA antibody titres of these fluids and serum assayedby ELISA-spot assay.RESULTSSpecific IgG was induced in RT and CE fluids at lowlevels (0.3-1.5% and 0.25-2.3% of the blood serumtitres respectively) following each immunisationregime, except following transdermal immunisationwhich induced higher levels (8.60/0 -RT fluid and7.6% -CE fluid) compared to the serum titre (Fig. I).The concentration of IgG in prostatic fluid followingrectal immunisation was higher than that induced atthe RT and CE and independent of the serum IgGtitre. The entry of 19A into the male fluids was alsofound to be independent of the blood 19A titre (Fig.2). 19A was detected in prostatic fluid following eachimmunisation regime, the highest titre being inducedfollo:ving IN immunisation at levels 3-8 fold higherthan In serum. 19A was present at low levels in fluidfrom t?e RT. fo.llowing each immunisation exceptrectal ImmUnISatIOn and only detected in CE fluidfollowing systemic immunisation. Specific 19A wasnot detected in seminal vesicle fluid following eithersystemic or mucosal immunisation.DISCUSSIONThe results show that some IgG enters at the rete testis(1) and is concentrated within the epididymal lumen. Theluminal titres seem to be dependent on blood titres, butonly reach a small proportion of them. The entry ofspecific 19A into the male fluids is interpreted as beingthrough local production by resident lymphocytes andsubsequent SC-mediated transport into the duct lumen.Similarly, it is concluded that IgG in prostatic fluidfollowing rectal immunisation arises due to its localproduction. Thus mucosal immunisation(s) may proveuseful in order to elicit an appropriate immune responseto antigens in the male reproductive tract.25:i 20a 15eJOr-----------------..2.20/..S,2Y. ,.EllITtluidI!lCEtluid5lPfluid:3 w 100.9%"'YSlJb.Q IntranasalRectal Transcbmll195 ------._--- --.. Serum1)KI'1I1id----Figure 1: The concentration (EU/ml) of anti-IT IgG antibody inreproductive fluids of male mice following various immunisationregimes. Mean ± SEM for 5 animals per group. % - percentage ofserum IgG titre..mOlid1!5 POlidFigure 2: The concentration (EU/ml) of anti-IT 19A antibody inserum and reproductive fluids of male mice following variousimmunisation regimes. Mean ± SEM for 5-10 animals per group,*p