We have previously reported that exposure of monkey embryos to 13‐cis‐retinoic acid (cRA) results in thymic defects. In this study, we analyzed lymphocyte and antigen‐presenting cell populations at gestational days (GDs) 80–100 in the thymus, spleen, mesenteric lymph nodes, and gut‐associated lymphoid tissue following a teratogenic dosing regimen of cRA (2.5 and 5 mg/kg) at GD14–27. Tissue sections were immunostained for T‐cells (anti‐CD3), B‐cells (anti‐CD20), dendritic cells (p55), and major histocompatibility class II (anti‐HLA‐DR). Digital images of spleen sections were analyzed to obtain the relative area occupied by the cell subsets within the white pulp (WP). Compared with controls, the T‐cell dependent compartment of the spleen WP in specimens with perturbed thymic development (aplasia and severe hypoplasia) showed a reduction in size and proportion of CD3⁺ T cells. Our findings indicate that cRA‐induced thymic defects result in disrupted development of the splenic T‐cell dependent compartment.