(A) FACS analysis of PBMC at day 0 and PBMC-derived floating cells cultured for 6 days without TCM (RPMI, day 6) or with TCM (TCM, day 6). T cells were doubly stained for CD3 and CD8 to obtain the percentage of CD8⁺ T cells (CD3⁺/CD8⁺), CD4⁺ T cells (CD3⁺/CD8⁻), CD8⁺ NK cells (CD3⁻/CD8⁺), and non-T cells (CD3⁻/CD8⁻). Staining with anti-CD14 and anti-CD20 antibodies was used to identify monocyte and B cells, respectively (left side). Shown are representative examples from 2 patients (upper and lower panels, respectively). Similar results were also obtained with floating cells from other 3 AIDS-KS patients and 1 asymptomatic HIV⁺ individual analyzed by immunocytochemistry (data not shown). (B) Phenotypic characterization of adherent cells obtained from PBMC of an AIDS-KS patient cultured for 6 days with TCM and analyzed by immunohistochemistry (APAAP method). (a) A representative negative control (isotype-matched control Ab) of adherent cells counterstained by hematoxylin with the typical spindle morphology is shown (original magnification × 400). (b through d) Representative examples of staining for CD68 (b; original magnification × 400), CD14 (c; original magnification × 1,000), and VE-cadherin (d; original magnification × 400) are presented. Nearly 100% of the cells present in these fields showed a specific red staining. The average percentage of positive cells from different fields for all the patients analyzed is shown in Table 5.