Novel humanized anti-CD3 antibodies induce a predominantly immunoregulatory profile in human peripheral blood mononuclear cells.
Strategies to minimize the immunogenicity and toxicity of murine anti-CD3 antibodies (e.g. OKT3) are of special interest for organ transplantation and for the treatment of autoimmune diseases. In the present work, we have developed two humanized anti-CD3 antibodies. These molecules were shown to bind to human CD3, though less efficiently, and display less mitogenic activity than OKT3. These results prompted us to investigate whether this reduced mitogenic potential was associated with the development of anti-inflammatory properties. Indeed, in peripheral blood mononuclear cells (PBMCs), the humanized antibody versions induced a predominantly anti-inflammatory cytokine profile, in contrast with the pro-inflammatory profile induced by OKT3. Neither OKT3 nor the humanized versions induced the expression of IL-4, IL-2 or TGF-beta. Both humanized antibodies induced significantly lower production of IFN-gamma and IL-5 and slightly higher production of IL-10 than OKT3. This immunomodulatory profile was most evident by the 80-fold higher ratio of IL-10/IFN-gamma production in PBMCs cultured in the presence of the humanized antibodies, compared to those stimulated with OKT3. Furthermore, these humanized anti-CD3 antibodies induced a late FOXP3 gene expression while OKT3 led to a more transient expression of FOXP3. Taken our results, we suggest that these humanized anti-CD3 antibodies may promote the development of T cells with immunoregulatory activity.
Silva
Hernandez M
HM
Departamento de Biologia Celular, Universidade de Brasília, 70910-900 Brasília, DF, Brazil.
Vieira
Pedro M M M
PM
Costa
Patricia L N
PL
Pimentel
Bárbara M S
BM
Moro
Ana M
AM
Kalil
Jorge
J
Maranhão
Andrea Q
AQ
Coelho
Verônica
V
Brigido
Marcelo M
MM
D000818:Animals; D000911:Antibodies, Monoclonal; D001327:Autoimmune Diseases / Q000276:immunology / Q000628:therapy*; D017252:CD3 Complex / Q000276:immunology*; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D016207:Cytokines / Q000378:metabolism; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D005818:Genetic Engineering; D006084:Graft Rejection / Q000276:immunology / Q000628:therapy*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics; D007167:Immunotherapy; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D016853:Muromonab-CD3 / Q000494:pharmacology; D016377:Organ Transplantation; D011993:Recombinant Fusion Proteins; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000473:pathology
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D016207:Cytokines; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D007128:Immunoglobulin Fragments; D016853:Muromonab-CD3; D011993:Recombinant Fusion Proteins