Effects of isotype and Fc region on in vitro function of a mouse/human chimeric CD4 antibody.
Murine CD4 mAbs have shown potential for the treatment of allograft rejection and autoimmune disorders including rheumatoid arthritis. Clinical usefulness of the murine mAbs has been limited by immunogenicity and a short circulating half-life. Mouse/human chimeric antibodies have been constructed, composed of the variable region of M-T412 (a murine G2a mAb specific for the human CD4 molecule) and human G1 (cM-T412 G1) or G4 (cM-T412 G4) Fc regions. F(ab')2 and F(ab) fragments of the murine G2a and chimeric G1 mAbs were generated by enzymatic digestion. The chimeric mAbs and all fragments retained the avidity and specificity of the murine M-T412 and were evaluated in in vitro assays measuring Ig production by pokeweed mitogen (PWM)-stimulated peripheral blood mononuclear cells (PBMC), sIL-2R produced by phytohemagglutinin-stimulated PBMC, and proliferation in response to tetanus toxoid, CD3 mAb plus IL-2, and mixed lymphocyte response (MLR). When PBMC were stimulated with tetanus toxoid, 10 ng/ml of cM-T412 G1 inhibited proliferation by 90%, while neither the cM-T412 G4, M-T412 G2a, nor any mAb fragment produced > 65% inhibition, even at 1000-fold higher concentrations. A similar pattern of inhibition was observed in MLR assays. In contrast, the F(ab')2 fragment of the cM-T412 G1 was as effective as the whole antibody in inhibiting PWM-stimulated IgM synthesis and PBMC proliferation in response to stimulation by a CD3 mAb plus IL-2.(ABSTRACT TRUNCATED AT 250 WORDS)
Dalesandro
M R
MR
Centocor Inc., Malvern, PA 19355.
Pak
K Y
KY
Tam
S
S
Wilson
E
E
Looney
J E
JE
Knight
D M
DM
Ghrayeb
J
J
Reiter
C
C
Rieber
E P
EP
Riethmuller
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D015704:CD4 Antigens / Q000378:metabolism; D006801:Humans; D007141:Immunoglobulin Fc Fragments; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M / Q000096:biosynthesis; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D051379:Mice; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D007141:Immunoglobulin Fc Fragments; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins