From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned...From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned into pGEM-T vectors, then tranfected into JM109. The VH and VL genes were snalyzed by automatic DNA sequencer. According to Kabat classification, the VH and VL genes belong to the mouse ig heavy subgroup Ⅱ(A) and x chain subgroupⅢ, respectively. The VH and VL genes were subcloned into pr1-Expr and Pk Expr respectively, then transfected into XL2-Blue. The VH- Pr1 and VL- pk were trans feeted by electroporation into mouse myeloma cell X63Ag8. 653. The transfectoma cells were selected by G418 screening, and then supernatant of cultured transfectoma were analyzed by ELISA and immunofluorescence techniques.We have acquired transfectoma cells secreting anti-CD4 chimeric antibodies.These chimeric antibodies are able to kill tumor cells specifically in vitro.展开更多
The precise mechanism underlying the effects of anti-CD4 antibody and calcium ions(Ca^(2+)) in peanut allergy remains unknown.C3 H/HeJ mice sensitized with peanut protein extract(PPE)were injected with anti-CD4 antibo...The precise mechanism underlying the effects of anti-CD4 antibody and calcium ions(Ca^(2+)) in peanut allergy remains unknown.C3 H/HeJ mice sensitized with peanut protein extract(PPE)were injected with anti-CD4 antibodies for 4 weeks.Stimulation with PPE increased the specific immunoglobulin E(IgE),cytokine,histamine,and mMcp-1 levels,upregulated decorin(Dcn)expression,induced Ca^(2+) inflow in the spleen,and augmented the expression of the transcription factors GATA-3 and Foxp3,which resulted in Th2 and Treg cell activation.Notably,the Ca^(2+) levels were positively correlated with the histamine,interleukin(IL)-4,IL-5,and IL-13 levels,and negatively correlated with IL-10 levels.However,administration of anti-CD4 antibodies markedly alleviated allergic symptoms,activated T cells,and reduced Ca^(2+) inflow,cytokine,histamine,mMcp-1,and the IgHG3,CXCLI2,MMP2 and FABP4 gene.Our results indicated that anti-CD4 antibodies can ameliorate PPE-induced allergy,which is probably related to the suppression of Ca^(2+) inflow,and inhibiting histamine,cytokine and IgHG3,CXCL12,MMP2,and FABP4,thus exerting a protective effect against PPEsensitized food allergy.展开更多
Tests show the monoclonal anti CD4 antibody (mAb) MT310 recognizes the gp120 binding site on CD4 as part of its mechanism for strongly inhibiting human immunodeficiency virus type 1 (HIV 1) infection of CD4 + T ...Tests show the monoclonal anti CD4 antibody (mAb) MT310 recognizes the gp120 binding site on CD4 as part of its mechanism for strongly inhibiting human immunodeficiency virus type 1 (HIV 1) infection of CD4 + T cells. In competition tests, mAb MT310 and mAb Leu3a (an anti CD4 mAb recognizing the gp120 binding site) all inhibited gp120 binding to CD4 + T lymphocytes, while mAb MT405 did not. This result suggests that MT310, like Leu3a, recognizes the gp120 binding site on CD4. To further confirm whether MT310 recognizes the gp120 binding site on CD4, we prepared rabbit anti idiotypic antisera (Ab2) against MT310 (Ab1). The anti idiotypic antisera against MT310 inhibited binding of MT310 and Leu3a to human CD4 + T lymphocytes, but did not block binding of MT151 with the second domain of CD4, while rabbit anti idiotypic antisera to MT151 could block binding of itself to these cells, but could not inhibit the binding of MT310 and Leu3a, further indicating that MT310 recognized the gp120 binding site on CD4.展开更多
文摘From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned into pGEM-T vectors, then tranfected into JM109. The VH and VL genes were snalyzed by automatic DNA sequencer. According to Kabat classification, the VH and VL genes belong to the mouse ig heavy subgroup Ⅱ(A) and x chain subgroupⅢ, respectively. The VH and VL genes were subcloned into pr1-Expr and Pk Expr respectively, then transfected into XL2-Blue. The VH- Pr1 and VL- pk were trans feeted by electroporation into mouse myeloma cell X63Ag8. 653. The transfectoma cells were selected by G418 screening, and then supernatant of cultured transfectoma were analyzed by ELISA and immunofluorescence techniques.We have acquired transfectoma cells secreting anti-CD4 chimeric antibodies.These chimeric antibodies are able to kill tumor cells specifically in vitro.
基金supported by the National Natural Science Foundation of China(31972185)。
文摘The precise mechanism underlying the effects of anti-CD4 antibody and calcium ions(Ca^(2+)) in peanut allergy remains unknown.C3 H/HeJ mice sensitized with peanut protein extract(PPE)were injected with anti-CD4 antibodies for 4 weeks.Stimulation with PPE increased the specific immunoglobulin E(IgE),cytokine,histamine,and mMcp-1 levels,upregulated decorin(Dcn)expression,induced Ca^(2+) inflow in the spleen,and augmented the expression of the transcription factors GATA-3 and Foxp3,which resulted in Th2 and Treg cell activation.Notably,the Ca^(2+) levels were positively correlated with the histamine,interleukin(IL)-4,IL-5,and IL-13 levels,and negatively correlated with IL-10 levels.However,administration of anti-CD4 antibodies markedly alleviated allergic symptoms,activated T cells,and reduced Ca^(2+) inflow,cytokine,histamine,mMcp-1,and the IgHG3,CXCLI2,MMP2 and FABP4 gene.Our results indicated that anti-CD4 antibodies can ameliorate PPE-induced allergy,which is probably related to the suppression of Ca^(2+) inflow,and inhibiting histamine,cytokine and IgHG3,CXCL12,MMP2,and FABP4,thus exerting a protective effect against PPEsensitized food allergy.
基金Munich University in Germany and theNational Key Basic Research Specific Funds of China(No.G19990 75 6 0 7)
文摘Tests show the monoclonal anti CD4 antibody (mAb) MT310 recognizes the gp120 binding site on CD4 as part of its mechanism for strongly inhibiting human immunodeficiency virus type 1 (HIV 1) infection of CD4 + T cells. In competition tests, mAb MT310 and mAb Leu3a (an anti CD4 mAb recognizing the gp120 binding site) all inhibited gp120 binding to CD4 + T lymphocytes, while mAb MT405 did not. This result suggests that MT310, like Leu3a, recognizes the gp120 binding site on CD4. To further confirm whether MT310 recognizes the gp120 binding site on CD4, we prepared rabbit anti idiotypic antisera (Ab2) against MT310 (Ab1). The anti idiotypic antisera against MT310 inhibited binding of MT310 and Leu3a to human CD4 + T lymphocytes, but did not block binding of MT151 with the second domain of CD4, while rabbit anti idiotypic antisera to MT151 could block binding of itself to these cells, but could not inhibit the binding of MT310 and Leu3a, further indicating that MT310 recognized the gp120 binding site on CD4.
