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Lymphoid and non-lymphoid subpopulations were investigated in the lung of the mouse with immunocyto-, immunohisto- and enzyme-histochemical methods. Special attention was paid to the cell populations in bronchus-associated lymphoid tissue (BALT), which is positioned between a bronchus and an artery. In BALT, discrete T- and B-cell areas can be found. The majority of the T cells belong to the L3T4+ (T-helper) subpopulation. In the T-cell area interdigitating cells can be recognized by anti-class II antibodies as well as by specific monoclonal antibodies, NLDC-145 and MIDC-8. Macrophage subpopulations can he discriminated by location, enzyme reactivity and various macrophage-specific monoclonal antibody markers. On the outer rim of BALT macrophages are recognized by the MOMA-1 and ERTR9 antibody. Macrophages dispersed in BALT can only be discriminated with the MOMA-2 antibody. The macrophage markers F4/80 and and Mac-1 show no reactivity in BALT. In lung, tissue macrophages around bronchi and blood vessels are predominantly recognized by the MOMA-1 and MOMA-2 antibody, and a minor population by the ERTR9 antibody. Alveolar macrophages show heterogeneity with the MOMA-1, MOMA-2 and NLDC-145 antibody. The relationship between alveolar macrophages and antigen-presenting cells is discussed here. [ABSTRACT FROM AUTHOR]

To investigate the role of the NLDC-145 antigenic determinant on interdigitating cells in T-cell dependent areas of lymphoid organs, rosettes were isolated from interdigitating cells and lymphocytes from lymph nodes. By using allotypic mixtures it was established that these rosettes represented pre-existing associations of interdigitating cells and lymphocytes. Phenotypic analysis of the lymphocytes showed that most cells in these rosettes were T cells. After antigenic stimulation, an increase of helper T cells and of B cells associated with the rosettes was found. Incubation of the rosettes with the NLDC-145 antibody did not alter the composition of the rosettes. By neonatal injection of the NLDC-145 antibody, the influence of the antigen on T-cell differentiation and antigen presenting was investigated. Although the antibody could be detected easily on interdigitating cells and thymus cortex of injected animals, no direct influence of long-term injections of the antibody from birth on antigen presentation and T-cell differentiation was found. [ABSTRACT FROM AUTHOR]

A single systemic dose of cyclophosphamide (CY) has been shown to enhance cellular immunity in a variety of antigen models. The immunoenhancing effects of CY have been attributed to its ability to selectively abrogate suppressor cell function. Previous studies from our group have demonstrated that local administration of distinct cytostatic drugs at the sensitization site can induce a similar enhancement of delayed-type hypersensitivity as systemic CY, with the obvious advantage of avoiding systemic side-effects. In the present study we investigated the effects of local administration of an optimally immunopotentiating dose of the active CY-derivative Z 7557 and, in selected experiments, of etoposide (VP- 16) and systemic CY on mononuclear cells in draining lymph nodes. Whereas CY caused a long-lasting and marked depletion of B-cell areas, locally administered Z 7557 and VP- 16 relatively spared B cells and even induced an increase in B- and 1-cell numbers in (keyhole limpet haemocyanin-) sensitized mice. At Day 4 the CD4JCD8 ratio was slightly reduced in drug- treated mice. Interestingly, drug treatment reduced the proportion of interdigitating cells staining with the monoclonal antibodies NLDC-145 and MIDC-8. Upon isolation, dendritic cells (DC) from sensitized, Z 7557-treated mice showed longer dendritic protrusions and an enhanced accessory cell function compared to DC from saline-treated controls. These findings suggest that immunoenhancing effects of cytostatic drugs may occur via an effect on DC. [ABSTRACT FROM AUTHOR]

A new monoclonal antibody, MIDC-8, is described which shows a comparable tissue distribution as the recently described NLDC-145 antibody. It recognizes interdigitating cells, veiled cells and Langerhans cells in lymphoid organs and the skin of the mouse. In contrast to NLDC-145 it recognizes a cytoplasmic component of these cell types. Its distribution is more restricted to the T cell-dependent areas than NLDC-145. Isolation of dendritic cells from lymph nodes, spleen and thymus revealed that both antibodies react with the in vitro isolated dendritic cells. The results show that these antibodies can be used to study dendritic cells in vitro and emphasize the relationship between the in vivo interdigitating cells of the T cell areas and the in vitro isolated dendritic cells.

Dendritic cells play an important role as accessory cells in the immunological defense system. Investigations into the biological functions of this population have been limited by the difficulties encountered in isolating sufficient numbers of cells, and therefore different approaches have been described to immortalize accessory cell populations. In this report, four murine hybrid cell lines (H1-H4) are described that were produced by fusion of in vitro isolated dendritic cells and a tumor cell. The hybrid lines are characterized by the expression of the dendritic cell specific marker NLDC-145, Ia expression and the absence of macrophage or T cell markers. The cell lines have no phagocytic activity and are functionally active as accessory cells. The expression of the NLDC-145 antigen in vitro was found to be inducible. These NLDC-145-positive cell lines will be useful for the further characterization of the NLDC-145 antigen and in the study of dendritic cell and lymphocyte interactions.

To investigate the role of the NLDC-145 antigenic determinant on interdigitating cells in T-cell dependent areas of lymphoid organs, rosettes were isolated from interdigitating cells and lymphocytes from lymph nodes. By using allotypic mixtures it was established that these rosettes represented pre-existing associations of interdigitating cells and lymphocytes. Phenotypic analysis of the lymphocytes showed that most cells in these rosettes were T cells. After antigenic stimulation, an increase of helper T cells and of B cells associated with the rosettes was found. Incubation of the rosettes with the NLDC-145 antibody did not alter the composition of the rosettes. By neonatal injection of the NLDC-145 antibody, the influence of the antigen on T-cell differentiation and antigen presenting was investigated. Although the antibody could be detected easily on interdigitating cells and thymus cortex of injected animals, no direct influence of long-term injections of the antibody from birth on antigen presentation and T-cell differentiation was found.

A monoclonal antibody, H1F5, is described that reacts with a subset of Lyt-2 (CD8) mouse T cells and LPS-activated B cells. In both lymph nodes and spleen of BALB/c mice, the H1F5 antigen is coexpressed approximately on 20%-30% of the CD8+ T cells and approximately on 91% of LPS-activated B cells. In the thymus, few cells (less than 1%) are positive for the marker, but no correlation could be demonstrated with markers for mature T cells such as MEL-14 and PNA expression. Elimination of H1F5+ cells by complement lysis led to a 30%-50% reduction of specific lysis as measured in a primary allo CTL, indicating that the cytotoxic effector cells are injured. The relationship of this marker and other antigenic determinants on lymphocytes is discussed.