Circadian and/or circahemidian rhythms in nine lymphocyte-related variables from peripheral blood of healthy subjects.

Circadian variations were investigated for nine lymphocyte-related variables in the peripheral blood of healthy subjects. Monoclonal antibodies targeted at membrane immunoglobulins (anti-Ig, anti-kappa, anti-lambda) or differentiation antigens (anti-IA and OKT3) were used to characterize respectively mature B cells (SIg+, kappa +, lambda +), cells expressing HLA-DR antigen (IA+), and T cells (OKT3+). Blood (33 ml) was drawn every 4 hr for 24 hr starting at 8.30 hr, on seven occasions in five apparently healthy male volunteers, recumbent from 23.00 hr to 07.00 hr. Leukocyte and differential counts were measured. Mononuclear cells were isolated on Ficoll-Hypaque before being incubated with monoclonal antibodies. The proportion of fluorescent cells per 100 microscopically determined cells was multiplied by the number of circulating lymphocytes per milliliter of venous blood. Temporal variations were validated by both paired t-test and cosinor. Rhythms with a period (tau) identical to 24 hr were validated with statistical significance (p less than 0.05) for total lymphocytes, OKT3+ cells and OKT3+:SIg+ ratio, and suggested (0.05 less than or equal to p less than or equal to 0.10) for lambda + and (kappa + + lambda +) cells. Rhythms with tau identical to 12 hr were also found (p less than 0.05) for OKT3+, SIg+, kappa +, and IA+ cells as well as for the OKT3+:SIg+ and the kappa +:lambda + ratios. Validated rhythms exhibited a large amplitude, e.g., peak-through differences were 40% of the 24-hr mean. This circadian and circahemidian temporal structure of immunologic variables constitutes a time-qualified reference system for investigating immune regulations and a tool for optimizing both diagnostic criteria and effectiveness of immunotherapeutic attempts.

A histological assessment of the four-day subrenal capsule assay (SRCA).

A four-day subrenal capsule assay (SRCA) was developed, since fragments of human tumours implanted under the renal capsule of immunocompetent mice became rejected by the host within six days. The assay requires a histological assessment of both its exploitability and the extent of drug-induced anti-tumour lesions. 45 tumours from 43 patients with solid tumour were submitted to an SRCA in 1410 male B6D2F1 mice. After being biopsied each tumour was dissected by a pathologist, cut into 50 pieces (1.5 mm3), and one piece was implanted under the renal capsule of 35 mice; the mean tumour diameter was measured on day 0. The mice were randomized into groups of 6 to 10 animals each. On days 1, 2 and 3, the mice were treated with either placebo (control group) or with various anticancer agents. On day 4 the animals were sacrificed, the mean tumour diameter measured, the tumour bearing kidney fixed in Bouin's picroformol solution and processed for histological analysis after staining with hematein. Fragments of fresh explants of human tumours retained their proliferative and metabolic capacity: mitoses were observed as well as keratinizing cells in epidermoid carcinomas and melanin-producing cells in melanomas. Proliferation of tumour cells was seen along the renal capsule suggesting their affinity for connective tissue. Capillaries filled with mouse erythrocytes were also seen. No or minimal lymphocytic infiltration was found. Drug oncolytic effects ranged from minor cellular degeneration to almost complete necrosis and were documented by the scoring of histologic lesions.(ABSTRACT TRUNCATED AT 250 WORDS)

A four-day subrenal capsule assay for testing the effectiveness of anticancer drugs against human tumors.

The subrenal capsule assay may predict to which anticancer drug a given patient's tumor is sensitive and may also be used to screen new anticancer drugs. The present study documents that the use of this model requires a histological assessment of both the exploitability of a subrenal capsule assay and the extent of drug-induced antitumor lesions. Thirty-five tumors from 34 patients with solid tumor were submitted to a subrenal capsule assay in a total of 1130 male B6D2F1 mice. After being biopsied, each tumor was dissected by a pathologist and cut into 50 pieces (1.5 X 1.5 X 1.5 cu mm), and one piece was implanted under the renal capsule of 35 mice; the mean tumor diameter was measured on Day 0. Mice were randomized into groups of 6 to 10 animals each. On Days 1, 2, and 3, mice were treated either with placebo (control group) or with various anticancer agents. On Days 4 or 6, mice were sacrificed, the mean tumor diameter measured, and the tumor-bearing kidney fixed in Bouin's picroformol solution and processed for histological analysis after staining with hematein -eosin. Seven histological parameters were blindly rated in a semiquantitative fashion yielding a compound score ( PAPAN ) which estimated the overall quality of each xenograft between -3 and +11. On Day 4, as opposed to Day 6, mean lymphocytic infiltration was 3-fold lower (p less than 0.01), and the rate of xenografts containing well-preserved cancer cells was 2-fold larger (p less than 0.01) in three different tumor specimens. Twenty-two of 31 (71%) assays were evaluable, as defined by a histological quality control test. In those, drug effects were demonstrable by statistically significant differences among groups in 2 assays (9%) by using the relative variation in tumor size as an index of drug effectiveness and in 12 assays (54%) by PAPAN histological score. This suggests the higher sensitivity of histological scoring over tumor size measurements. Moreover, no correlation between relative variation in tumor size and PAPAN was demonstrable with statistical significance indicating the poor reliability of tumor size measurements as an index of the antitumor effectiveness of cytostatic drugs.