Anti-CD4 Recombinant Antibody (Clenoliximab)

A, Keliximab-mediated adhesion of IFN-treated monocytic cell line (THP1) to CD4 fibroblast transfectants. Also shown is the inhibition of this CD4 mAb-mediated adhesion by sCD4 along with the effect of truncated form of keliximab. B, Comparison of IgG1 and IgG4 anti-CD4 mAbs in FcR-CD4-mediated cell-cell adhesion assay. Percent adhesion of THP-1 cells to CD4 T cells was calculated as: (fluorescence of test samples/total fluorescence) 100. Data points are the mean SEM of triplicate cultures.

Reddy, M. P., Kinney, C. A. S., Chaikin, M. A., Payne, A., Fishman-Lobell, J., Tsui, P.,... & Reff, M. (2000). Elimination of Fc receptor-dependent effector functions of a modified IgG4 monoclonal antibody to human CD4. The Journal of Immunology, 164(4), 1925-1933.

A, Keliximab and clenoliximab-mediated adhesion of CD4 T cells to IFN-induced Fc receptor-expressing monocytes. Anti-CD4 mAbs serve as a bridge between the monocytes on the microtiter plates and fluorescent dye loaded SupT1-18 T cells. Percent binding of SupT1-18 cells to monocytes was calculated as in Fig. 4B (fluorescence of test samples/total fluorescence) 100. Data represent one of three experiments that showed identical results (15). B, Freshly isolated human monocytes were cultured for 36 h with 250 U/ml human IFN-, after which induction of Fc receptor expression was measured by staining with anti-human CD64-FITC, CD32-FITC, and CD16-FITC Abs. Flow cytometric analysis was performed using CellQuest software (Becton Dickinson).

Reddy, M. P., Kinney, C. A. S., Chaikin, M. A., Payne, A., Fishman-Lobell, J., Tsui, P.,... & Reff, M. (2000). Elimination of Fc receptor-dependent effector functions of a modified IgG4 monoclonal antibody to human CD4. The Journal of Immunology, 164(4), 1925-1933.

Shown are flow cytometric two-dimensional scatter plots of human PBMCs incubated with 100 ng/ml of keliximab or clenoliximab or medium without added Ab and dual stained for CD4 (OKT4) and CD3 markers after 24 h of culture.

Reddy, M. P., Kinney, C. A. S., Chaikin, M. A., Payne, A., Fishman-Lobell, J., Tsui, P.,... & Reff, M. (2000). Elimination of Fc receptor-dependent effector functions of a modified IgG4 monoclonal antibody to human CD4. The Journal of Immunology, 164(4), 1925-1933.

IL-2, IFN-g, IL-4, and IL-10 were measured in serum by ELISA and plotted as picograms per milliliter. The results were obtained from pooled spleens of three to five mice, and the values shown are the mean 6 1 SD from three or four separate experiments. Supernatants were analyzed from cells treated with medium (f) or stimulated with Ad-lacZ (M). These analyses were performed with muCD4KO, HuCD4, or HuCD4 treated with Clenoliximab (CD4 mAb). p, p, 0.001, using Student's t test.

Chirmule, N., Truneh, A., Haecker, S. E., Tazelaar, J., Gao, G. P., Raper, S. E.,... & Wilson, J. M. (1999). Repeated administration of adenoviral vectors in lungs of human CD4 transgenic mice treated with a nondepleting CD4 antibody. The Journal of Immunology, 163(1), 448-455.

This figure summarizes the T cell proliferative and humoral responses in HuCD4 mice treated with multiple doses of vector in the presence or the absence of Clenoliximab. A and B, Splenocytes obtained on various days were analyzed for adenovirus- and SEB-induced lymphoproliferative responses as described in Materials and Methods. Data are presented as the stimulation index, which is the incorporation of radioactivity in the presence of stimulator (i.e., adenovirus or SEB) over the incorporation of isotope when cells are incubated with medium alone. Analyses were performed on days 28, 60, and 90. A, Adenovirus-induced lymphoproliferation (LPR) responses; B, SEB-induced LPR responses. C and D, The humoral immune responses of animals that received multiple administrations of vector in the presence or the absence of Clenoliximab. C, Neutralizing Ab from serum; D, neutralizing Ab from BAL (both presented as reciprocal dilutions).

Chirmule, N., Truneh, A., Haecker, S. E., Tazelaar, J., Gao, G. P., Raper, S. E.,... & Wilson, J. M. (1999). Repeated administration of adenoviral vectors in lungs of human CD4 transgenic mice treated with a nondepleting CD4 antibody. The Journal of Immunology, 163(1), 448-455.

Animals were euthanized on days 4, 28, 60, and 90 (rows 1, 2, 3, and 4, respectively) and analyzed for expression of b-galactosidase by X-galactosidase histochemistry. Magni-
fication is 3200. The results are representative of one of six mice in each group.

Chirmule, N., Truneh, A., Haecker, S. E., Tazelaar, J., Gao, G. P., Raper, S. E.,... & Wilson, J. M. (1999). Repeated administration of adenoviral vectors in lungs of human CD4 transgenic mice treated with a nondepleting CD4 antibody. The Journal of Immunology, 163(1), 448-455.