Vol 27 No 2 December - The Indian Society for Parasitology

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88 CSF production induced by L. donovani amastigote components JPD : Vol. 27 (2), 2003 assay medium consisted of 0.8% methylcellulose, static. The LDAA-induced augmented serum CSF 30% FBS, 10% pokeweed mitogen (Sigma)- production was observed as early as 6 h after stimulated spleen cell-CM, 2 mM glutamine and 5 x administration, reached its maximum after 24 h, and -5 10 M 2-ME. The spleen and BM cells were then gradually returned to normal levels within 72 h. resuspended in CFU-GM assay medium and plated in Control antigen and HI-LDAA (10 mg/kg each), and 5 5 sterile 35 mm petri dishes at 2 x 10 and 3 x 10 only,DMEM did not induce the production of CSFs. cells/dish, respectively. The number of CFU-GM was LDAA-induced CSF production by macrophages, determined according to colony morphology after 7 in vitro: LDAA (0.01-1 mg/ml), in a concentrationdays of incubation in humidified 5% CO2-air dependent manner, induced CSF elaboration by atmosphere at 37°C. Based on the total spleen or BM -4 -6 elicited peritoneal MØs (3x10 -10 cells/dish; Table cell numbers, the final CFU-GM counts were I). Whereas, a low concentration of 0.01 mg/ml LDAA expressed/spleen or femur. could induce detectable levels of CSFs in the CM, the Statistical analysis: All experiments were run in maximum response (73 ± 9 colonies) occurred at 0.1 triplicate, three-times, separately. Student's t-test was mg/ml; at higher concentrations the response became used for statistical analysis, and p
JPD : Vol. 27 (2), 2003 CSF production induced by L. donovani amastigote components 89 elaborated CSFs (Table II); here again 0.1 mg/ml LDAA induced maximum CSFs (69 ± 10 and 77 ± 10 colonies by splenic and BM-derived MØs, respectively) after 24 h. Furthermore, LDAA (0.01-1 mg/ml) did not directly induce the colony formation (data not shown). Neutralization of LDAA with polyclonal rabbit anti-LDAA antibody significantly (p
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Vol 27 No 2 December - The Indian Society for Parasitology

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