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Cell surface marker evaluation of infant Macaca monkey leukocytes in peripheral whole blood using simultaneous dual‐color immunophenotypic analysis

Abstract: Cross‐reactivity between several commercially available mouse antihuman monoclonal antibodies (mAbs), conjugated to phycoerythrin (PE) and fluorescein isothiocyanate (FITC) fluorochromes, and peripheral blood leukocyte surface antigens, has been established in infant cynomolgus (Macaca fascicularis) monkeys using whole blood lysis, and two‐color, PE and FITC flow cytometric analysis. With the exception of the CD8 marker, the bivariate dot‐plot patterns for all other markers were similar in infant monkeys and in humans. For the CD8 marker, however, a CD8+CD2− population of cells was observed in the majority of monkeys tested (10 out of 12). The number of CD8+CD2− cells was higher (13%) in infant monkeys compared to the 3% reported for adult human blood. The mean percentage and absolute numbers for the cell surface markers identified with the human mAbs CD2 (FITC, Ortho, Paritan, NJ), CD4 (PE, B‐D, Mountain View, CA), and CD8 (PE, B‐D) when these were combined with a series of PE‐ or FITC‐labelled human mAbs were similar across all combinations tested. Statistically significant differences were observed between male and female monkeys for the mean percentage levels of CD4 (females > males) and for the CD4/CD8 ratio (females > males). Such gender differences need to be taken into consideration when infant cynomolgus monkeys are used as models for chemical‐induced immunotoxicity studies. Measurement of the interleukin‐2 (IL‐2) and transferrin proved to be useful in monitoring in vitro cellular activation in infant cynomolgus and possibly in rhesus (M. mulatta) monkeys.