Two methods were used to demonstrate the presence of tumor-specific antigens in adenocarcinomata of the human colon: (a) rabbits were immunized with extracts of pooled colonic carcinomata, and the antitumor antisera thus produced were absorbed with a pooled extract of normal human colon and with human blood components; (b) newborn rabbits were made immunologically tolerant to normal colonic tissue at birth, and were then immunized with pooled tumor material in adult life. Normal and tumor tissues were obtained from the same human donors in order to avoid misinterpretation of results due to individual-specific antigenic differences. The antisera prepared by both methods were tested against normal and tumor antigens by the techniques of agar gel diffusion, immunoelectrophoresis, hemagglutination, PCA, and immunofluorescence. Distinct antibody activity directed against at least two qualitatively tumor-specific antigens, or antigenic determinants, was detected in the antisera prepared by both methods and at least two additional tumor antigens were detected exclusively in antisera prepared by the tolerance technique. Whether these additional antigens were qualitatively different from normal tissue antigens, or merely present in tumor tissue in higher concentrations than in normal tissue has not as yet been determined. Furthermore, it was shown that the tumor-specific antibodies were not directed against bacterial contaminants or against the unusually high concentrations of fibrin found in many neoplastic tissues. It was concluded from these results that the pooled tumor extracts contained tumor-specific antigens not present in normal colonic tissue. Identical tumor-specific antigens were also demonstrated in a number of individual colonic carcinomata obtained from different human donors.

https://rupress.org/jem/article-pdf/121/3/439/1267132/439.pdf

Demonstration of tumor-specific antigens in human colonic carcinomata by immunological tolerance and absorption techniques

Staphylococcal enterotoxins and a group of related proteins made by Streptococci cause food poisoning and shock in man and animals. These proteins share an ability to bind to human and mouse major histocompatibility complex proteins. The complex ligand so formed has specificity for a particular part of T cell receptors, V beta, and by engaging V beta can stimulate many T cells. It is likely that some or all of the pathological effects of these toxins are caused by their ability to activate quickly so many T cells. It is also possible that encounters with such toxins have caused mice, at least, to evolve mechanisms for varying their T cell V beta repertoires, such that they are less susceptible to attack by the toxins.

https://science.sciencemag.org/content/sci/248/4956/705.full.pdf

The staphylococcal enterotoxins and their relatives

1) Induction of humoral antibody formation involves the obligatory recognition of two determinants on an antigen, one by the receptor antibody of the antigen-sensitive cell and the other by carrier antibody (associative interaction). 2) Paralysis of antibody formation involves the obligatory recognition of only one determinant by the receptor antibody of the antigen-sensitive cell; that is, a nonimmunogenic molecule (a hapten) can paralyze antigen-sensitive cells. 3) There is competition between paralysis and induction at the level of the antigen-sensitive cell. 4) The mechanisms of low- and high-zone paralysis, and maintenance of the unresponsive state, are identical. 5) High-zone paralysis occurs when both the carrier antibody and the receptor antibody are saturated, so that associated interactions cannot take place. 6) The mechanisms of paralysis and induction for the carrier-antigen-sensitive cell are identical to those for the humoral-antigen-sensitive cell. 7) The formation of carrier-antigen-sensitive cells is thymus-dependent, whereas humoral-antigen-sensitive cells are derived from bone marrow. Since carrier antibody is required for induction, all antigens are thymus-dependent. 8) The interaction of antigen with the receptor antibody on an antigen-sensitive cell results in a conformational change in an invariant region of the receptor and consequently paralyzes the cell. As the receptor is probably identical to the induced antibody, all antibody molecules are expected to be able to undergo a conformational change on binding a hapten. The obligatory associated recognition by way of carrier antibody (inductive signal) involves a conformational change in the carrier antibody, leading to a second signal to the antigen-sensitive cell. 9) The foregoing requirements provide an explanation for self-nonself discrimination. Tolerance to self-antigens involves a specific deletion in the activity of both the humoral- and the carrier-antigen-sensitive cells.

https://science.sciencemag.org/content/sci/169/3950/1042.full.pdf

A Theory of Self- Nonself Discrimination.

A series of monoclonal antibodies was used to define three discrete stages of human intrathymic T-cell differentiation The earliest stage was confined to <10% of thymocytes, which were·reactive with both OKT9 and OKT10 Subsequently, approximately 70% of human thymocytes acquired a thymocyte-restricted antigen, OKT6, lost OKT9 antigen, and expressed reactivity with OKT4 and OKT5 These last two monoclonal antibodies were previously shown to define inducer (helper) and cytotoxic/suppressor populations, respectively, in peripheral blood The OKT4+, OKT5+, OKT6+ “common” thymocyte population represents the majority of thymocytes and accounts for more than 70% of thymocytes With further maturation, thymocytes lose OKT6 reactivity, segregate into OKT4+ and OKT5+ subsets, and acquire reactivity with OKT3 (and OKT1) This latter stage corresponds to the more functionally mature subset The possible relationship of acute lymphoblastic leukemia of T-cell lineage to these proposed stages of intrathymic differentiation was determined Analysis of 25 tumor populations showed that 21 could be related to one or another differentiative stage The majority (15/21) were derived from an early thymocyte or prothymocyte subpopulation, 5/25 were derived from a common thymocyte subpopulation, and 1/25 was derived from a mature (OKT3+) subpopulation These data suggest that is it now possible to define stages of T-cell differentiation that can be related to T-cell malignancies in humans

https://www.pnas.org/content/pnas/77/3/1588.full.pdf

Discrete stages of human intrathymic differentiation: Analysis of normal thymocytes and leukemic lymphoblasts of T-cell lineage

The differentiation and function of human T lymphocytes

The effects of Salmonella endotoxin and staphylococcal enterotoxin B (SEB) were examined in vitro on mouse and human lymphocyte populations from unsensitized subjects. These substances were found to be extremely stimulatory to mouse spleen cells as measured by 3 H-thymidine incorporation. In regards to size, general morphologic features, and staining properties, the cell cultures appear identical to cultures stimulated by phytohemagglutinin (PHA). However, endotoxin failed to stimulate human peripheral lymphocytes while PHA and SEB produced a response over a wide dosage range. Attempts to detect stimulation of human lymphoid cells by endotoxin using various mammalian sera as augmentors were unsuccessful.

The mitogenic effects of endotoxin and staphylococcal enterotoxin b on mouse spleen cells and human peripheral lymphocytes.

IN VIEW Of the success of splenectomy in dealing with certain clinical problems, and the lack of any clearly unfavorable effects of this procedure, the idea that the spleen is not physiologically essential has been widely accepted. The first serious challenge to this concept arose with the clinical observation of King and Schumaker of a high incidence of serious bacterial infection following splenectomy performed in infancy. This original report has been supplemented during the last decade by additional clinical data, and renewed interest in the role of the spleen in immunity has resulted. Recent experimental data have defined more clearly immunological functions for the spleen that are of special significance during early development. The purposes of this are paper are (1) to review the clinical problem which led to re-examination of the role of the spleen in immunity, particularly, as it relates to the early development of the immune mechanism, and (2) to offer a hypothesis which may explain the clinical observation of overwhelming infection after splenectomy in the young infant. THE CLINICAL PROBLEM In 1952 King and Schumaker reported an unusual incidence of infection in infants who had undergone splenectomy for congenital hemolytic anemia. The hazards of splenectomy in early life were pointed out in a number of papers which appeared subsequently with large series of cases reinforcing the validity of the earlier observations. Most recently Horan and Colbatch have incisively reviewed the problem and added a number of their own cases to those already in the literature. In spite of the preponderance of evidence, dissenting reports have appeared and the subject remains somewhat controversial.

The role of the spleen in immunity. With special reference to the post-splenectomy problem in infants.

Two cases of congestive cardiomyopathy were proved to be due to chronic atrial automatic-focus tachycardia. Surgical elimination of the tachycardia resulted in normalization of cardiac function in both cases. We suggest that patients with congestive cardiomyopathy be carefully screened for supraventricular tachycardia. Patients with chronic supraventricular tachycardia should be treated aggressively to prevent myocardial dysfunction. ( JAMA 1985;253:391-392)

https://jamanetwork.com/HttpHandlers/ArticlePdfHandler.ashx?journal=jama&articleId=396342&pdfFileName=jama_253_3_028.pdf

Chronic supraventricular tachycardia. A curable cause of congestive cardiomyopathy.

Membrane Immunofluorescence Reactions of Burkitt Lymphoma Cells From Biopsy Specimens and Tissue Cultures

The media and culture conditions required for in vitro stimulation of mouse lymphoid cells are described. The medium was arginine-rich and contained heat-inactivated human serum. A component of the human sera necessary for stimulation of the cells was a natural mouse cell agglutinin, which affected both background stimulation and the degree of induced stimulation with phytohemagglutinin (PHA). Absorption of the agglutinin from the human serum rendered the medium incapable of sustaining DNA synthesis in the presence of PHA.

The response to PHA of mouse spleen and thymus cells was age-dependent and, although this response was not present at birth, it rapidly rose to adult levels. Spleen cells from mice immunized with bacillus Calmette-Guerin (BCG) or sheep erythrocytes (SRBC) showed increased in vitro reactivity to added purified protein derivative (PPD) or SRBC stroma, dependent on the time of immunization.

The dose response curve for the SRBC stroma stimulated, immune spleen cells is compatible with a theory of cell to cell interaction being necessary for an in vitro reaction to antigen.

The possible role of the mouse cell agglutinin (AMLG) is discussed.

/pdf/cellular-recognition-by-mouse-lymphocytes-in-vitro-i-4ag0dd08o8.pdf

Cellular recognition by mouse lymphocytes in vitro i. definition of a new technique and results of stimulation by phytohemagglutinin and specific antigens

Richard T. Smith

Papers

The mitogenic effects of endotoxin and staphylococcal enterotoxin b on mouse spleen cells and human peripheral lymphocytes.

The role of the spleen in immunity. With special reference to the post-splenectomy problem in infants.

Chronic supraventricular tachycardia. A curable cause of congestive cardiomyopathy.

Membrane Immunofluorescence Reactions of Burkitt Lymphoma Cells From Biopsy Specimens and Tissue Cultures

Cellular recognition by mouse lymphocytes in vitro i. definition of a new technique and results of stimulation by phytohemagglutinin and specific antigens