Human peripheral blood phagocytes ingest Escherichia coli 026:B6 lipopolysaccharide (LPS)-coated paraffin oil droplets containing Oil red O only if fresh serum deposits C3 on the surfaces of the particles (opsonizes them), by reactions involving the properdin system. The rate of binding of purified [125-I]C3 in serum to LPS-coated particles correlated precisely with the rate of acquisition of ingestibility assayed spectrophotometrically. Once opsonized, LPS-coated particles remained fully ingestible and retained fixed [125-I]C3 radioactivity even after exposure to extremes of temperature, pH, ionic strength, phospholipases, urea or guanidine, some nonionic and ionic detergents, and organic solvents. Trypsin, human conglutinogen-activating factor, another heat-stable activity found in human serum, and sodium dodecyl sulfate removed radioactivity and diminished ingestibility of the opsonized particles. Alkylation, reduction plus alkylation and F(ab')2 from anti-C3 blocked ingestibility but did not alter particle-bound radioactivitymelectrophoretic and tryptic peptide autoradiographic analysis of dodecyl sulfate eluates of opsonized particles, cleansed of many contaminating proteins by boiling with 2 M NaCl (yet still opsonized), revealed that the polypeptide with C3-derived radioactivity had a mol wt of approximately 140,000 and was composed of 70,000 mol wt subunits linked by disulfide bonds. Immunochemical analysis and comparison of the peptide structure of the eluate with that of C3 indicated that the opsonic fragment is not the fragment defined as C3b but a smaller derivative of C3.
Antibody responses to pneumococcal polysaccharide vaccine were compared in a control group of 17 normal adults and in a group of 27 adult patients with stable renal function (serum creatinine 0.8--2.1 mg/dl) seven months to nine years following renal transplantation. Using the indirect hemagglutination technique, antibody titers to 13 of the 14 capsular antigens contained in the vaccine were determined for each patient just prior to and again three weeks following immunization. There was no significant difference between the two groups in the proportion of patients responding with a fourfold rise in titer to 12 of the 13 antigens tested. The response rate to antigen type 3 was reduced in the transplant group (p less than 0.05). Mean fold increase in indirect hemagglutination titers was likewise determined for each antigen, and a reduced response in the transplant group was noted only to antigen type 23 (p = 0.037). Immunosuppressed renal allograft recipients appear capable of mounting a nearly normal antibody response to pneumococcal vaccine.
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