Erythrocytes were prepared by washing with isotonic sodium iodide to elute adsorbed serum proteins and then resuspended in five minutes BSA/HBSS to your concentration of 2 108/ml. A level of 200 l of FITC labeled bacteria was incubated with 10 l of NHS, alone or along with different percentages of MAb to form 3 capsule, at 37 C for 30 min while shaking. Afterwards, 200 m of erythrocytes was added and the incubation was continued for 30 min. After washing with 0. 1% BSA/HBSS to get rid of unbound bacteria, the adherent bacteria c-Met inhibitor and erythrocytes were set with 1% paraformaldehyde for flow cytometry. Erythrocytes were gated, and 20,000 events were mentioned. The MF of erythrocytes was calculated for each test. To measure the erythrocyte adherence mediated by human anti supplement antibody, bacteria were incubated with 10 l of normal mouse serum as a typical supply of complement, alone or along with 10 l of heatinactivated human pre or postvaccination serum. Erythrocyte adherence was determined by subtracting the erythrocyte adherence exhibited in normal mouse serum from that exhibited in normal mouse serum plus pre or postvaccination serum. Exchange reaction experiments were done exactly as in the erythrocyte Infectious causes of cancer adherence analysis described above, except that after the bacteria were washed from the erythrocytes, 200 m of J774A. 1 macrophages was added and the mixture was incubated at 37 C for 30 min while shaking. The erythrocytes were then lysed with BD FACS lysing solution for 10 min at room temperature. After washing with 0. 1% BSA/HBSS, the macrophages were fixed with 1% paraformaldehyde and analyzed by flow cytometry. Macrophages were gated, and 15,000 activities were collected. The MF of macrophages was used to measure the exchange reaction. The normal fluorescence natural product library of macrophages was taken from each test. To gauge the involvement of CR3 and Fc RIII/II in mediating the transfer response, the macrophages were preincubated with rat anti mouse CD11b or rat anti mouse CD16/CD32 MAb at 37 C for 30 min, after which the macrophages were washed and put into the erythrocytes as described above. The transfer reaction was done with normal mouse serum as a standard source of complement, alone or along with heatinactivated human pre or postvaccination serum, to judge the transfer reaction mediated by human anti tablet antibody. To determine the effects of the mouse MAb to type 3 capsule on complement C3, C1q, and C4 deposit onto the pneumococcal surface, type 3 pneumococcal tension WU2 and its nonencapsulated mutant JD908 were opsonized in NHS alone or together with different concentrations of MAb to type 3 capsule. The floor bound C3, C1q, and C4 were then detected by flow cytometry. We found that in the lack of MAb to form 3 capsule, complement C3 deposition onto Cps3 stress WU2 was lower than that onto the Cps3 isogenic mutant JD908, though similar levels of C1q and C4 were placed on JD908 and WU2.