New CD4(+) cell line susceptible to infection by HIV-1.

Infection of a newly described human T lymphoid cell line, CEM-CL10, with three different variants of HIV-1 resulted in cytopathic effects followed by cell lysis. Following primary lytic infection, proviral DNA could not be detected by Southern blot analysis in the outgrowth of the surviving CEM-CL 10 cells 60 days after initial exposure to HIV-1. These surviving cells could be further grown as a separate line, derived from CEM-CL10, and were found to be resistant to subsequent infection by HIV-1. A marked decrease in CD4 antigen expression was observed in these latter cells but not of the CD3 and transferrin receptor antigens. This decline in cell surface CD4 expression was correlated with both an absence of specific CD4 mRNA and with changes in structure of the CD4 gene. Both the HIV-1-sensitive CEM-CL10 cell line and its CD4(-), HIV-1-resistant derivative line, will be made available to interested investigators.

Cellular composition of rat bone marrow stroma. Antigen-defined subpopulations.

Although stromal cells establish the architecture of mammalian bone marrow and organize hemopoiesis, the interrelationships among their macrophage, fibroblastic, endothelial, and adipocyte-like components are not wholly understood. Using murine monoclonal antibodies to cultured adherent cells of rat bone marrow, we observed that the predominant fibroblastoid cells grown from marrow differed from those of non-hemopoietic organs. The marrow type bore a detectable quantity of the ST3 but not ST4 antigen, whereas those from lung, diaphragm, and epididymal fat pad, bore more ST4 than ST3. Those from spleen were an equal mix of both types. Although the tissue distribution of the ST3 antigen was similar to that of Thy-1, it was not identical, and in the brain, the two structures were localized in different areas. While none of the ST3, ST4 (fibroblast directed), or BN(MB)35 (myeloid directed) antibodies recognized fat cells cultured from marrow, the ST10 antibody, selected for binding to marrow derived fat cells, stained peripheral adipose cells, unidentified aglobular cells in areas of fat cell formation, and macrophages, but not fibroblasts. On the basis of these observations, we suggest that the fibroblastoid cells of the marrow are different from those of non-hemopoietic tissues.

Deficiency of myeloperoxidase and abnormal chromosome 1 occurs in variant (HL60) promyelocytes.

Maturation of normal polymorphonuclear neutrophils is characterized by successive periods of granule synthesis, a process which frequently is abnormal in leukemia. Recently, the human leukemic cell line HL60, displaying a promyelocytic phenotype, has been used to study granulocyte maturation. We describe a variant line of HL60, called HL60-A7, resulting from growth in actinomycin D, which contains atypical large azurophilic granules deficient in myeloperoxidase. The products of in-vitro translation of A7 RNA contained less than 5% of the immunoreactive MPO found in the parent line. Electrophoresis of plasma membrane polypeptides radioiodinated by the lactoperoxidase technique revealed several differences. Karyotypic analysis identified a consistent chromosome 1q+ abnormality which was not found in any of the parental cells examined. This constellation of differences between HL60 and HL60-A7, i.e. MPO deficiency, abnormal granule morphology, cell surface changes, and further cytogenetic abnormalities, may point to a common site sensitive to altered regulation in some leukemic promyelocytes.

Antigens differentially expressed on surface and cytoplasmic structures of human myeloid cells.

Specialized internal granules are a major feature of myeloid differentiation and are deficient in most acute myeloid leukemia cells. Although they arise from the same synthetic apparatus as does the plasma membrane, their relationship to it is not well characterized in human tissues. Using murine monoclonal antibodies, we have identified myeloid-related structures that illustrate three possible modes of antigen expression in these organelles. Immunocytochemical studies with the light microscope have shown that the first (D51) was restricted to the surface of neutrophils, monocytes, megakaryocytes and platelets; a second (D46) was found on the surface of blastic cell lines but on only internal components of mature granulocytes; the third (H36/71) appeared on both the surface and internal particles of promyelocytes, myelocytes and polymorphs. These model antigens may be used to study the control of granule synthesis in normal and leukemic cells.

Quantitation of helper factor activity. I. Utilization of the ELISA method to measure helper activity in the secondary antibody response.

We describe here a simple culture system for measuring the effects of regulatory cells or factors on the secondary antibody response which uses the enzyme-linked immunosorbent assay (ELISA) to detect antibody production. This technique offers several advantages over methods which measure hemolytic plaque-forming cells. Helper factors purified from the supernatant of in vitro primed helper cells were shown to specifically enhance antibody production from primed spleen cells by from 2- to 4-fold in this system.