Freshly collected chronic lymphocytic leukemia B cells (B-CLLAPC. 11 Moreover, transfection of B7-1 (CD80) or B7-2 (CD86) cells) are known to be inefficient at stimulating allogeneic T into tumor cells strongly enhances antitumor T cell responses cells, and to lack significant expression of B7 (CD80 and CD86) in murine models. [12][13][14][15] Considering that normal resting B cells

SUMMARYWe investigated the phenotype and functional capacities of tumour-infiltrating lymphocytes (TIL), freshly isolated from primary renal cell carcinoma (RCC) specimens (n ¼ 20). Three-colour flow cytometry immunophenotyping revealed that RCC TIL consist mainly of CD3 þ T cells, with a clear predominance of CD4 ¹ CD8 þ over CD4 þ CD8 ¹ T cells, and a marked population of CD4 þ CD8 þ T cells. Natural killer (NK) cells were also strongly represented (> 25% in 15 of 20 tumour samples), while B cells constituted a minor TIL subset (< 5% in 18 of 20 tumour samples). More importantly, the T and NK cells within the tumour displayed a significantly higher expression of the early activation marker CD69 than their counterparts in adjacent normal renal tissue and in peripheral blood. Expression of CD54 and of HLA-DR was also elevated on CD3 þ TIL, and HLA-DR expression was further vigorously up-regulated following ex vivo stimulation with anti-CD3, all suggesting enhanced immune activity within the tumour microenvironment. CD3 þ CD4 þ TIL displayed a normal capacity to up-regulate CD25 expression and to secrete both Th1-type (IL-2, tumour necrosis factor-alpha (TNF-a) and interferon-gamma (IFN-g)) and Th2-type (IL-4, IL-5 and IL-10) cytokines upon triggering with anti-CD3. Furthermore, cytokine production was susceptible to modulation by CD28 costimulation. CD3 þ CD8 þ TIL, on the other hand, consistently demonstrated a poor up-regulation of CD25 upon triggering with anti-CD3, and displayed poor ex vivo cytolytic activity in an anti-CD3-redirected 4-h cytotoxicity assay against murine P815 cells. Collectively, our findings indicate that the CD3 þ CD4 þ TIL in RCC have normal functional capacities, whereas the proportionally major CD3 þ CD8 þ TIL are functionally impaired. The relevance of these findings to the in vivo local immune response in RCC is discussed.

No abstract

Three-color flow cytometry immunophenotyping revealed significant increases of CD57 + and CD28 − cells among both circulating CD4 + and CD8 + T lymphocytes of untreated hemato-oncological patients (n = 54) as compared to healthy donors (n = 55), with CD57 and CD28 expression on the patients' T cells being largely reciprocal. Marked expansion of CD57 + cells among circulating CD4 + T lymphocytes was frequently detected in patients with chronic leukemia of B cell origin (B-CLL, hairy cell leukemia) but not in patients with chronic myeloid leukemia, suggesting a causal relation with the tumor's major histocompatibility complex class II expression. Using immunomagnetic separation techniques, we further demonstrate that the patients' CD57 + /CD28 − T cells display a typical Th1-type cytokine secretion profile upon anti-CD3 stimulation, with a markedly higher secretion of the Th1-type cytokines IL-2, IFN-␥, and TNF-␣ than their CD57 − /CD28 + counterparts. Cytotoxic activity of circulating CD8 + T lymphocytes, measured ex vivo in an anti-CD3-redirected assay, was almost exclusively exerted by the CD57 + /CD28 − subset. Moreover, a marked cytotoxic activity was detected within CD4 + CD57 + T cells from some B-CLL patients. Finally, the patients' CD57 + /CD28 − T cells displayed an increased tendency to apoptosis in culture. Collectively, our results indicate that the expanded CD57 + /CD28 − T cells in hemato-oncological patients represent differentiated effector cells, similar to their (quantitatively minor) counterpart in healthy donors. The reason for their expansion and their pathophysiologic significance, however, remains unclear.

Advanced multivariate data‐analytical techniques are proposed to concisely represent and evaluate complex lymphocyte profiles (i.e., compound lymphocyte subset distributions) of individual subjects in easily interpretable, two‐dimensional, graphical correlation biplots. The lymphocyte profile of each subject is represented by its location in the model, and the score of a subject for a particular lymphocyte subset is inferred from the perpendicular projection on a rotated axis that coincides with this lymphocyte subset. Simultaneously, the model yields information about the correlation between the lymphocyte subsets. Furthermore, individuals with aberrant lymphocyte profiles can be easily identified. In case studies of 80 healthy donors and of 40 patients with multiple myeloma, 10 patients with monoclonal gammopathy of undetermined significance, and 50 age‐ and sex‐matched healthy donors, reconstruction of the two‐dimensional lymphocyte profiles from 27 flow‐cytometric characterized lymphocyte subsets succeeded in representing 43% and 51% of the total information (variability) contained within the 80 × 27 (= 2,160) and 100 × 27 (= 2,700) flow cytometry measurements, respectively. It is concluded from the present studies that the correlation biplot represents a unique and powerful tool to concisely describe, represent, and analyze complex lymphocyte profiles of individual subjects and the heterogeneity in lymphocyte profiles among these subjects. Cytometry 28:220–227, 1997. © 1997 Wiley‐Liss, Inc.