Gordon Watkins Douglas researched cervical cancer, breach delivery, and treatment of high blood pressure during pregnancy [5] in the US during the twentieth century. He worked primarily at Bellevue Hospital Center in New York, New York. While at Bellevue, he worked with William E. Studdiford to develop treatments for women who contracted infections as a result of illegal abortions performed throughout the US in unsterile environments. Douglas also established the first contraception [6] and pregnancy [5] termination clinic at Bellevue Hospital shortly after the legalization of abortion [7] as a result of the 1973 US Supreme Court ruling in Roe v. Wade [8]. Furthermore, Douglas showed that fetal and maternal cells exchange between the pregnant woman and fetus [9] during pregnancy [5], which led to the later development of non-invasive prenatal testing in the early twenty-first century.

Body odors regulate social, sexual and endocrine responses of many species. Specialized structures have evolved to produce and detect odorous signals. Individual identity, often communicated through body odor, can be critical in mate choice, incest avoidance, parental care and other inter-individual interactions. Because of the importance of individual identity, particularly among social species, we have focused our research program on understanding how odors code for individuality and in what contexts individual signals modulate social behavior. In particular, we have been investigating how one set of genes, those of the MHC (see below), are involved in producing an animal’s self odor.

Two-dimensional chymotryptic peptide maps of Lyt-1.1 and Lyt-1.2 from Lyt-1 congenic thymocytes labeled with 125I in the usual way before lysis did not significantly differ, but there was a characteristic difference between maps of Lyt-1.1 and Lyt-1.2 obtained from 125I-labeled solubilized membrane fragments. We conclude that the Lyt-1 locus of chromosome 19 includes the protein-structural gene for Lyt-1. This conclusion is further supported by evidence with 35S-cysteine-labeled thymocytes: Thus an early 62K intermediate form, and a 60K form from tunicamycin-treated cells devoid of N-linked and O-linked carbohydrates, were precipitated by Lyt-1 alloantibody, which implies that the allo-specificity of Lyt-1 glycoprotein (67K) resides partly or wholly in protein.

The major histocompatibility complex (MHC) imparts to each mouse an individual urinary odor, called "odortype", which reflects its MHC genotype. Perception of odortypes affects mate selection and embryonic implantation. Recent findings that odortypes are expressed as early as one day of age suggested that they might already be expressed in utero. We now report that at 9-12 days of gestation, odortypes specified by paternal (non-maternal) MHC haplotypes become apparent in maternal urine. Thus, odortypes are expressed in utero, can be sensed even before birth, and may serve in familial identification and communication.

Of various labels that have been attached to antibody, ferritin is the most useful visual marker for locating antigens at the high degree of resolution permitted by electron microscopy, and ferritin-labeled antibody has been used extensively to study the surface structure of viruses, bacteria, and cells of various sorts (1). The technique developed by Singer involves the coupling of ferritin to the antibody by metaxylylene diisocyanate or toluene 2,4-diisocyanate (2, 3). Alternatively, ferritin may be coupled to an anti-q,-globulin antibody, providing an "indirect" immunoferritin method based on the same principle as the indirect immunofluorescence test; in both instances there is the advantage that a single labeled antiserum suffices for the recognition of more than one antigen. Recently, Hiimmerling et al. (4) devised a new method of ferritin labeling that avoids undesirable consequences of coupling ferritin to antibody chemically. Hybrid F(ab')~ antibody is prepared according to Nisonoff and Rivers (5) from two sera, rabbit anti-mouse "l~G and rabbit anti-ferritin, each hybrid molecule having two different combining sites, one for mouse ~G and the other for ferritin. To locate, for instance, a cell surface alloantigen, the cells are first exposed to the relevant alloantibody (mouse ~'G), then to the hybrid antibody anti-mouse q~G/ anti-ferritin, and finally to ferritin. The new method has been used so fax only in this indirect form, although it is equally applicable in the direct form (e.g. with anti-alloantigen/antiferritin hybrid antibody). Its outstanding advantage is that it opens the way to the use of a variety of visual markers, such as viruses, the only requirements being that these be morphologically distinguishable, of suitable size, and immunogenic in the species in which the hybrid partner antibody is to be prepared. Exemplifying the application of immunoferritin methods to the study of cell surface structure, ABO group alloantigens have thus been found to cover all or most of the human erythrocyte, while Rh alloantigens are confined to limited areas (6, 7). Among nucleated cells, many surface antigens of mouse lymphoid cells, both normal

As the cells that are programmed to generate the various sets of immunocompetent lymphocytes progress along their several pathways they pass through discrete phases of differentiation that can be recognized by the selective expression of cell surface components. Detailed investigation of the nature and mechanisms of these differentiative steps has been much facilitated by the development of short-term assays in which some of these phenotypes are induced in vitro (1-4). We present here an account of our findings with induction assays that monitor an early step in T-cell differentiation, marked by acquisition of the surface component Thy-1, and two steps in B-cell differentiation, the first marked by acquisition of complement receptors (CR) 1 (5) and the second by acquisition of the surface component PC (plasma cell antigen) (6).

The alloantigen G,x is found on the surface of thymocytes of some strains of mice (G, x+), where it is demonstrable by complement (C)-dependent lysis (cytotoxicity assay) with (W/Fu x 13N)FI anti-W/Fu(C58NT)D (anti-NTD)' serum, and not on the thymocytes of other strains (G,x) . Its expression in mice of the 129 strain, where its genetics has been studied in most detail, is controlled by two unlinked chromosomal loci (1) . (Control of its expression in other G,x+ mouse strains may differ in detail from that of strain 129, and this is a subject of continuing study [2] .) An important feature of G, x antigen is its relation to leukemia virus and leukemogenesis . Thus, cells other than thymocytes, and from G, x as well as G, xmouse strains (and from rats), may express G, x antigen if they become productively infected with murine leukemia virus (MuLV) (1) . In the accompanying paper, Obata et al . (3) give serological evidence that G, x antigen is a constituent of the gp69/71 glycoprotein component (4, 5) of the MuLV envelope, implying that the mendelian inheritance of G, x in certain mouse strains signifies the mendelian inheritance of a component of MuLV expressed selectively on cells following the T-lymphocyte differentiative pathway. We report here a related study in which a molecule resembling gp69/71 was isolated from intact thymocytes of 129 mice by enzymatic radioiodination of the thymocyte surface, followed by release of this glycoprotein with the nonionic detergent Nonidet P-40 (NP-40) and precipitation with anti-NTD serum; the product was then compared with envelope proteins isolated from purified MuLV. Our interpretation is that G,x antigen is carried by a glycoprotein with a mol wt

I t is remarkable that of the six separate systems of surface antigens identified on thymocytes and lymphocytes of the mouse with cytotoxic antisera (1), all but one (H-2) are represented only, or mainly, on these cell types. Another antigen system with characteristically limited tissue representation has recently been recognized on plasma cells (2). We have referred to such discriminatory surface components as "differentiation antigens" (1). Thus the antigenic constitution of the surfaces of immunocompetent cells, and by inference also the spectrum of genes used by these cells to build their surface membranes (1), is characteristic of this pathway of differentiation. Continuing our study of the antigenic composition of cells belonging to the immune system, we have examined the question "What immunoglobulin components can be identified as surface elements of immunocompetent cells of different types?" There is already a variety of evidence from other laboratories that Ig components are in fact detectable on such cells (see review ref. 3). To do this we have studied the reactivity of a range of anti-Ig sera in tests with various immunocompetent cell populations, using the same cytotoxicity test system that has been used to identify all the other known surface antigens of immunocompetent cells referred to above. The Ig phenotypes of these cells were then further defined by testing the ability of various anti-Ig sera to interfere with their activity in a number of well-defined tests that measure functions of immunocompetent cells. In discussion, we shall at tempt to summarize present information on the surface phenotypes of different cells belonging to the various compartments of the immune system, including all the known differentiation antigens which serve to distinguish one such population from another, of which the Ig components represent one category.

A major achievement of cancer research in animals has been the demonstration that several tumors of natural occurrence are caused by viruses. One of the foremost goals of human cancer research is now to identify viruses that may be responsible for any form of cancer in man. Two groups of viruses have so far emerged as important in the natural etiology of animal tumors: (a) DNA viruses of herpes-type, responsible for the Luck~ adenocarcinoma of frogs (1) and Marek's neurolymphomatosis of chickens (2), and (b) the RNA oncogenic viruses (oncornaviruses0 responsible for leukemias and sarcomas of the chicken, mouse, and cat (~, 5). Among human cancers there is evidence associating a herpes-type virus with Burkitt's lymphoma and nasopharyngeal carcinoma, including reports that it may induce transformation of human leukocytes in vitro (6, 7). On the other hand, because of the many known examples of sarcomas and leukemias induced by oncornaviruses in animals, evidence has been sought for a similar causative virus in man. Virus of this type is often demonstrable by electron microscopy in sarcomas and leukemias of the chicken, mouse, and cat, but this has not been the case with comparable human tumors and leukemias. In man, particles with the morphology of oncornaviruses have only seldom been demonstrable and then usually in numbers so small as to raise questions about their significance (8). Nevertheless this is not strong evidence against participation of oncornaviruses in human malignancy, because many animat tumors which unquestionably are of viral origin, and which carry viral genomes, do not produce complete virus particles. For these reasons, the relatively negative morphological evidence is inconclusive.

Major histocompatibility complex (MHC) genes confer individual olfactory identity that can be detected with exquisite accuracy by mice. The fact that MHC genes themselves generate the characteristic odortype, rather than dedicated odor-determining genes, was supported in studies of point mutations in H2K and HLA transgenic mice, which evinced distinct odor profiles in olfactory assays. In this article we provide further evidence for a central role of MHC genes themselves in odortype specification by demonstrating that mice that are unable to express their genomic class I MHC genes because they lack beta2-microglobulin are distinguishable by scent from otherwise identical mice which possess an intact B2m gene. This odortype disparity appears at 9-12 days of gestational age, the period in which the MHC is first detectable in fetal cells of normal mice.

Genetically determined body odors that distinguish one mouse from another are termed odortypes. The best known odortypes, highly expressed in urine, are those specified by H-2, the major histocompatibility complex of the mouse, but other odortypes originate from unidentified loci in the rest of the genome, including both sex chromosomes. The definition of H-2 odortypes and evidence that their perception affects reproductive behavior have so far depended on studies with inbred mouse strains whose genetic differences are confined to the H-2 complex of genes. To simulate feral conditions more closely, a freely segregating population was bred from crosses involving four unrelated inbred strains contributing four different H-2 haplotypes. After H-2 typing, this outbred population was divided into four groups of freely segregating mice, comprising the four distinct H-2 genotypes represented, to serve as conventional donors of urine for evaluation in the standard Y-maze system used in the training and testing of mice for H-2 odortype discrimination. With respect to utility in training mice for H-2 odortype discrimination, and to degrees of concordance attained in the Y-maze by trained mice, these urinary H-2 odortype sources from outbred mice were no less effective than urines customarily obtained for those purposes from nonsegregating inbred donors. We conclude that discrimination of H-2 odortypes is not appreciably affected or impaired by the usual concurrent segregation within the genome as a whole.

Studies into the effects of aging on the immune system are hampered by the lack of a suitable animal model that is readily available and cost efficient. The mutant mouse, hairless (hr/hr genotype), has been shown to undergo an accelerated thymic involution with accompanying immunodeficiency. Thus, this strain of mouse has been proposed as a model for studying the interactions of aging and immune function. We have investigated the effects of homozygous hr gene expression over time on the immune function of these mice. It was observed that homozygous hr gene expression had minimal effects on peripheral lymphocyte subset compositions but did appear to result in changes in thymic differentiation. Further, hr/hr mice displayed decreased proliferative responses to IL2 and mitogen stimulation, although cytotoxic responses (both NK and T cell mediated) appeared normal. These defects appear to be attributable to T helper cell dysfunction. Each of the changes found in hr/hr mice were distinct from those seen with age-matched control mice. Thus, the hr/hr inbred strain of mouse does not appear to be a suitable model for use in analyzing the effects of aging on the immune system.

The major histocompatibility complex (MHC) is the prime but not exclusive determinant of genetically specific constitutive body odors, termed odor types, represented strongly in urine of the mouse. Perception of MHC-determined odor types influences reproductive behavior in the contexts of mate choice and maintenance of early pregnancy, tending to favor the propagation of one MHC type over another. How MHC genotype determines MHC odor type is unknown. One possible explanation is that differential odorants are generated by populations of commensal microorganisms whose composition is somehow geared to MHC diversity. This hypothesis was tested in the Y-maze system in which mice are trained to distinguish the urinary odors of MHC-congenic mice. First, it was shown that mice could readily be trained to distinguish the urines of germfree MHC-congenic mice. Second, it was shown that mice trained to distinguish the urines of conventionally maintained MHC-congenic mice could as readily distinguish the urines of germfree MHC-congenic mice. These results imply that MHC-determined odor types do not depend on odorants generated by microorganisms.

Ly-5 (CD45) glycoproteins of the mouse, expressed by all or most hematopoietic cell lineages and specified by a single Ly-5 gene, range in size from isoform T200 of T cells (the smallest), in which exons 4, 5, and 6 are not represented, to isoform B220 of B cells (the largest), in which all three of these optional exons are represented. The main purpose of the present study, utilizing the polymerase chain reaction (PCR), was to ascertain whether known isoforms of intermediate size are generated by single or dual usage of optional exons 4, 5, and 6. Transcripts representing all eight isoforms predictable from varied use of three exons were observed among a diverse panel of nine B-cell tumors in culture, but there was no evident concordance with known contrasting differential features that distinguish members of the B-cell tumor panel. No two B tumors exhibited the same variety of transcripts and the relative quantities of transcripts expressed varied greatly from tumor to tumor. Cloning of B-cell tumors did not alter their distinctive transcript patterns. Separation methods (sodium dodecyl sulfate polyacrylamide gel electrophoresis; SDS-PAGE) did not suffice to segregate all corresponding expressed isoforms but did establish that transcripts representing usage of a single optional exon and of two optional exons were actually translated, which supports a provisional inference that all eight isoforms exist. The considerable diversity of B-cell transcript phenotypes was not seen among seven T-cell leukemias, two cytolytic T-cell lines, and three Th1 helper T-cell lines, all of which displayed a uniform phenotype comprising major expression of the T200 transcript (no optional exon) and minor expression of a transcript employing exon 5. However, a panel of five cloned Th2 T-cell lines, which represents a second and functionality different branch of the helper/inducer T-cell category, exhibited a characteristic transcript pattern which distinguished them from a panel of three Th1 T-cell lines. The major transcript in the Th2 lines was also T200, but the Th2 lines showed higher representation of transcript containing optional exons. A single Th2 clone expressed an unusual transcript suggesting a potential isoform not compounded simply by varied inclusion of the three identified optional exons. After activation of the helper T-cell lines with concanavalin A (Con A), expression of transcript containing optional exons appeared to decrease.