T cell subsets and cellular immunity in end-stage renal disease.

The T lymphocyte population was studied by immunofluorescent staining with monoclonal antibodies and laser flow cytometry in the blood of 50 patients with end-stage renal disease undergoing long-term maintenance intermittent hemodialysis. The absolute number of T cells was lower in patients receiving dialysis for more than one year (p less than 0.001), as was the absolute count of helper T cells (p less than 0.005). In patients under 30 years of age, the absolute number of helper T cells was markedly reduced, whereas the number of suppressor/cytotoxic T lymphocytes was not changed. In patients between the ages of 30 and 60 years, both helper and suppressor cells were significantly reduced. In patients over 60 years of age, only the number of helper T cells was reduced. The in vitro response of patients' lymphocytes was reduced both in the mixed lymphocyte reaction (p less than 0.01) and after phytohemagglutinin stimulation (p less than 0.001). Natural killer cytotoxicity of patients' peripheral blood mononuclear cells, however, was unaffected.

NuMA: evaluation of a new biomarker for the detection of low stage colorectal cancer.

An enzyme immunoassay for NuMA was evaluated in a retrospective clinical study for its potential utility in the detection of colorectal cancer. The concentrations of NuMA and CEA (Abbott IMx) were measured in sera from 86 patients (presurgical) with colorectal cancer, 72 subjects with benign gastrointestinal diseases, 80 subjects with risk factors for colorectal cancer, and 141 age-matched healthy subjects. Reference values for NuMA and CEA were calculated by two methods: 95% cumulative distribution and ROC analyses versus healthy subjects. By the first method, NuMA and CEA both had approximately 20% sensitivity for colorectal cancer. By the second method (which generated lower reference values), NuMA was more sensitive than CEA for colorectal cancer. This improved sensitivity was most evident in Dukes B subjects. By either analysis method, NuMA was more sensitive than CEA for subjects at risk for developing colorectal cancer, whereas CEA was more specific for benign gastrointestinal diseases.

Methodologies for enumerating peripheral T lymphocytes.

Flow cytometric or manual microscopic techniques can be employed for the identification and quantitation of T lymphocytes in peripheral blood. Two flow cytometric devices (the Ortho Spectrum III and the Cytofluorograf FC200) were compared with each other and with two manual methods, each of the latter performed by two independent investigators, to evaluate the technical sensitivity and reproducibility of each method. Flow cytometry proved to be the most sensitive and consistent procedure for the enumeration of T cells. Among the manual methods examined, the enhanced immunofluorescence technique was found to be the least subjective and most reproducible.

Subpopulations of chicken B lymphocytes.

Immunoglobulin-synthesizing cells from the spleen and bursa were fractionated by the 1 X G sedimentation velocity technique and characterized by their ability to synthesize immunoglobulin and by staining with fluorescent anti-light chain chain. Four subpopulations of immunoglobulin-synthesizing cells were identified. In the bursa, slowly sedimenting (S 2.3 mm/hr) and rapidly sedimenting (S greater than 3.5 mm/hr) subpopulations with surface immunoglobulin were present; in the spleen, a slowly sedimenting (S 2.3 mm/hr) subpopulation with surface immunoglobulin and plasma cells (S greater than 3.5 mm/hr) with large concentrations of intracellular immunoglobulin existed. The subpopulations differed most markedly in their ability to synthesize immunoglobulin (cpm Ig synthesized/10(6) Ig-positive cells); the rates of immunoglobulin synthesis were in the ratio 1:2:1:900. The slowly sedimenting B cells from the spleen and both subpopulations of B cells from the bursa released small amounts of immunoglobulin into the culture media, whereas, the plasma cells released immunoglobulin at a rate as much as 3700 times greater. Bursal B cells could be further distinguished from splenic B cells by a greater amount of DNA synthesis.

Functionally distinct stages in B-lymphocyte maturation.

Evidence is reviewed that supports the concept of extrinsic stem cell colonization of inductive micro-environments for B-cell formation in the bursa of Fabricius of chickens and foetal liver of mammals. In these organs, the first distinctive marker of B-lineage differentiation (readily detectable immunoglobulin) is synthesized and displayed on cells which have extensive proliferative and differentiative capacity. The degree of commitment of these cells to the synthesis of trace or large quantities of immunoglobulin of the respective major classes when they colonize peripheral lymphoid tissues and the role of antigen in this process are considered.

Creating a useful panel of anti-T cell monoclonal antibodies.

Monoclonal antibodies to human T lymphocyte surface markers were produced by cell fusions between splenocytes from mice, immunized with T lineage cells, and mouse myeloma cells. Our approaches to immunization, clone selection and analysis of the resultant monoclonal antibodies with similar reactivities were produced. In one example, we found that several distinct monoclonal antibodies identified the same T inducer subset yet, apparently, recognized epitopes of the differentiation antigen(s) on these cells.

Evaluation of E-rosetting human lymphocytes with OKT11 and other monoclonal antibodies.

Monoclonal antibody OKT11 was found to compete with sheep red blood cells for binding sites on human lymphocytes. Preincubation of lymphocytes with OKT11 eliminated E-rosette formation. In a study of 142 peripheral blood samples ranging from 1% to over 90% E-rosette-positive cells, comparison to the percent OKT11-positive cells yielded a correlation coefficient of 0.93. In normal donors, subsets of OKT11+ cells were identified using two-color immunofluorescent staining methods with OKT3, OKT4, and OKT8. On the average, approximately 13% of OKT11+ lymphocytes were OKT3- and 13% of OKT11+ lymphocytes were OKT4- and OKT8-. Based on our double antibody fluorescence intensity data, low antigen density OKT11+ lymphocytes were OKT3-. OKT4+ and OKT8+ lymphocytes in normal peripheral lymphocytes have similar OKT11 antigen density.