Toxic shock syndrome toxin-1 (TSST-1) induces IL-1 production from macrophages/monocytes of various species. Susceptibility of the mouse strains to TSST-1 is parallel to the IL-1 production. IL-1 has multibiological activities for T cells, hepatocytes, neutrophils, fibroblasts and other cells, resulting in induction of acute phase responses. PI 5 IL-1 (IL-1 alpha) and PI 7 (IL-1 beta) are cloned in both the human and the mouse cases (1_??_3). The recombinant IL-1's confirm the multiple biological properties of the purified IL-1 including fever. IFN-alpha is also an endogenous pyrogen. Recombinant tumor necrosis factor (TNF) possesses almost the same biological function of IL-1. However, TNF induces IL-1 production from human mononuclear cells while IL-1 production by IL-1 itself has never been reported, yet. Differently from IL-1, TNF does not augment T cell proliferation. Although TSST-1 induces only IL-1, endotoxin stimulates both IL-1 and TNF production. A quantitative relationship among these endogenous pyrogens remains unsolved in each disease.
1) Auron, et al.: Proc. Natl. Acad. Sci. USA, 81: 7907, 1984.
2) Lomedico, et al.: Nature, 312: 458, 1984.
3) Gubler, et al.: J. Immunol, 136: 2492, 1986.

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We studied the effect of interleukin-3 (IL-3) on colony formation by hemopoietic progenitors in methylcellulose cultures of spleen cells from 5-fluorouracil (FU)-treated mice. Purified IL-3 supported the growth of various types of multilineage colonies including blast cell colonies. IL-3 supported the growth of multilineage colonies from single cells isolated from blast cell colonies by micromanipulation. This results shows that IL-3 acts directly on multipotential progenitors. Analysis of colonies derived from paired progenitors revealed disperate lineage expression and was in accordance with the stochastic model of stem cell differentiation.
Next, we examined the effect of purified IL-3 and erythropoietin on colony formation in serum-free cultures. In the presence of IL-3 alone, most of the multilineage (three or more lineages) colonies did not contain erythroid cells. However, in the presence of IL-3 and erythropoietin, most of the multilineage colonies contained various numbers of erythroid cells. Replating experiments suggest that IL-3 maintains the growth of the progenitor cells, which could differentiate into erythroid cells. Erythropoietin facilitated the terminal differentiation and amplification of erythroid cells, although it did not sustain the growth of multipotential stem cells. Single cell transfer experiments demonstrate that IL-3 supported the late stages of differentiation of neutrophils, macrophages, and mega karyocytes in the absence of lineage specific factors.
IL-3 plays a permissive role in proliferation and differentiation of multilineage hemopoietic progenitors in culture, and does not determine the differentiation pathway of cells.

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A cell surface marker analysis is one of the most useful tool for diagnosis and monitoring of the hematological diseases. Single and two color analyses of lymphoid disorders using monoclonal antibodies and a flow cytometer (FACS 420) were discribed here. The single color analysis has become to be popular at many labolatories and its usefulness is widely recognized. Recently the two color analysis with a flow cytometry has become available and a more precise analysis is expected.

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Modern topics on immunophenotypic and immunogenotypic analysis of lymphoproliferative disorders are summarized.
Recently developed various monoclonal antibodies against lymphocyte cell surface molecules have made it possible to classify B and T cells into functionally distinct detailed subsets.
In these days, in addition to histopathological study, analysis of cell surface markers (immunophenotypic analysis) of lymphocytic leukemias and lymphomas comes to be necessary information to diagnose, detect the prognosis, and select the treatment protocols.
Moreover, it has been revealed that many of the surface markers themselves serve as functional molecules to generate the subset specific functions. For example, T3 molecule, pan T cell marker, is a T cell antigen receptor (TCR)-related structure, and considered to be closely associated to T cell recognition and activation. Precise phenotypic analysis revealed that both T3 and interleukin 2 receptor (IL2R) are abnormally expressed on adult T cell leukemia (ATL) cells. ATL cells might be a useful tool to investigate the role of T3 molecule in T cell activation. Phenotypic analysis may not only classify the diseases, but also offer reseach materials of the special conditions of lymphocytes.
On the otherhands, it is now available to follow the process of gene rearrangements of immunoglobulin (Ig) and TCR in lymphoid cells, using appropriate probes of each gene segments and Southern transfer technique. Such DNA analysis (immunogenotypic analysis) is now extensively carried out in many institutions. By these means, it comes to be possible to determine clonality and cell lineage of the lymphocyte population.
Because the first step of Ig and TCR gene rearrangement occurs before lineage specific surface molecules appear, it is possible to determine the cell lineage of phenotypically nonT nonB lymphoid tumors. Not a small population of nonT nonB type were found to have Ig and TCR genes both in rearranged states.
Here, two important knowledges brought from the clinical studies are introduced.
One is that the study of each chain of Ig gene rearrangement in lymphoid tumors has determined the hierarchy of each chain rearrangement within a cell. Rearrangement occurs in the order of Heavy, s and finally λ chain.
Another is to bring to light the relation of chromosomal abnormalities frequently seen in lymphoid neoplasmas and the locus of Ig, TCR gene and oncogenes. In many cases, at chromosomal break point, Ig or TCR gene and an oncogene, such as c-myc, sit side by side, and the expression of the oncogene is increased.
Finally, one example of phenotypic and genotypic analysis of T cell lymphocytosis is indicated.

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The percentages of the peripheral blood lymphocytes from 156 healthy individuals at different ages (20_??_98 yr olds) of both sexes and 11 patients with Werner's syndrome (premature aging syndrome) were studied by the use of monoclonal antibodies and two-dimensional flowcytometry. Pan-T monoclonal antibodies positive cells (Leu-1⁺ or Leu-4⁺) were decreased with age, which was attributable to a relative decrease in the Leu-2a⁺suppressor/cytotoxic T cells, more precisely in the Leu-2a⁺ Leu-15^- cytotoxic T cells. In contrast to the cytotoxic T cells, Leu-3a⁺ Leu-8^- helper T cells and Leu-4⁺ HLA-DR⁺ activated T cells were increased in proportion with age in the healthy controls. These age-related changes of the lymphocyte subpopulations were also observed in some patientswith Werner's syndrome.

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