Human IgG3 is decreased and IgG1, IgG2 and IgG4 are unchanged in molecular size by mild reduction and reoxidation without any major change in effector functions.
Purified proteins of the four human IgG subclasses were reduced under neutral conditions to break the interchain S-S bonds, followed by dialysis to allow reformation of S-S bonds (pr/o treatment). The IgG1, IgG2 and IgG4 proteins apparently reformed native molecules by pr/o treatment, while IgG3 formed molecules with significantly smaller size, as measured by HPLC gel filtration, compared to the autologous native proteins. The degree of shrinking of the pr/o IgG3 molecules varied and was most pronounced at low protein concn. In addition, the temp and the concn of reducing agent during the pr/o treatment had some influence on the molecular size. The effect is probably due to a conformational change of the 62 amino acid long hinge of IgG3. The effector activity of pr/o IgG2 and pr/o IgG3 was studied by employing chimeric, mouse V and human C regions, monoclonal antibodies with the same NIP-binding properties. Thus, the interaction between IgG and the complement system was unchanged both for pr/o IgG2 and pr/o IgG3, while the Fc-receptor-mediated antibody-dependent cellular cytotoxicity (ADCC) was depressed to the same degree for both pr/o IgG2 and pr/o IgG3. Conclusively, the alteration of the conformation of the IgG3 molecule by pr/o treatment had no major influence on its effector functions.
Michaelsen
T E
TE
Department of Immunology, National Institute of Public Health, Oslo, Norway.
Naess
L M
LM
Aase
A
A
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003167:Complement Activation / Q000276:immunology; D004229:Dithiothreitol / Q000494:pharmacology; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006358:Hot Temperature; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D007132:Immunoglobulin Isotypes / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D008968:Molecular Conformation; D008970:Molecular Weight; D010084:Oxidation-Reduction; D011486:Protein C / Q000494:pharmacology
D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D011486:Protein C; D004229:Dithiothreitol