bluuebunny/crossref_metadata_2025 huggingface dataset:
A Study of the Laboratory Aids to the Diagnosis of Chronic Gonorrhea in Women
Authors: Patterson Raymond S.
Published: December 2022

In acute gonococcus infections a clinical diagnosis usually may be confirmed by a microscopical examination of a stained smear, but in chronic gonorrhea no single laboratory test can be relied upon to confirm the clinical diagnosis. Torrey, Wilson, and Buckell (1) studied the available laboratory procedures and conclude that whenever possible the smear, cultural, and complement-fixation tests should all be carried out as they tend to supplement each other; and that the value of the several tests increases in the order named, the complement-fixation test being most useful.

The conclusion of these investigators confirms the premise of this paper that there is no single, satisfactory test available for the confirmation of the diagnosis of chronic gonorrhea in women. As the control of the spread of gonorrhea is dependent upon a satisfactory test to detect gonorrhea in persons presumed to be infecting others, the need prompted a reconsideration of the value of the accepted laboratory procedures and a study of an additional diagnostic method which hitherto has not been reported as applied to the diagnosis of gonococcus infections.
Heparin Inhibition of Anaphylactic Shock
Authors: Kyes Preston , Strauser E. R.
Published: December 2022

A decrease in blood coagulability in anaphylactic shock has been described by Biedl and Kraus (1), Friedberger (2) and others for the guinea pig, rabbit, and dog. Gahringer, however, in the current number of this Journal shows that the change of primary importance as to blood coagulability, is an increase in coagulability concurrent with the acute symptoms of shock and that the decreased coagulability is a negative phase secondary to this increase. An extension of Gahringer's work in this laboratory shows also that the same sequence of changes in coagulability obtains in a great variety of shock reactions induced by the injection of water-insoluble particles such as bacteria, colloidal carmine, lamp black, etc. It would appear then, that during the occurrence of acute anaphylactic and anaphylactoid shock symptoms, there is an increased tendency on the part of the fibrinoplastic substances to undergo those changes which lead to the formation of fibrin aggregates.
On the Cold Agglutinins in Human Serum
Authors: Landsteiner K. , Levine Philip
Published: December 2022

The existence of two qualitatively different subtypes in Group II bloods is confirmed with the method of cold agglutination. Two cold agglutinins are shown to be present in Group II sera, one of these acting on bloods AA1, the other on bloods AA2. Group I bloods are characterized by the presence of a structure which is not common to all bloods, but which seems to be similar to that of A2. Some observations made with cold agglutinins would indicate that there exist a number of individual differences in human bloods, the phenomena being much less pronounced than typical isoagglutination. From several known facts it seems probable that the individual differences such as have been demonstrated by serological reactions are due to variations in substances other than proteins.
The Inflammatory Response and Resistance to Endotoxin in Mice
Authors: Moeller G. Radford , Terry Linda , Snyderman Ralph
Published: December 2022

Endotoxin lipopolysaccharide is a potent inflammatory agent which produces many pathophysiologic effects in mice. One strain of mice, C3H/Hej, is unique in its responses to endotoxin in that, in contrast to other mice, it possesses a high degree of resistance to endotoxin lethality, does not support an in vitro mitogenic response to endotoxin, and produces a small immune response to endotoxin in vivo. To see if the in vivo inflammatory responses produced by endotoxin in the resistant and normally susceptible mice were consistent with the above dichotomies, we investigated the kinetics of the i.p. inflammatory response to endotoxin in C3H/Hej mice vs normally susceptible strains. Paradoxically, the C3H/Hej mice responded with a rapid i.p. influx of PMN which peaked at 12 hr. This was followed by an influx of macrophages which peaked at 48 hr. In susceptible strains, the maximum PMN and macrophage responses were at 96 hr and were at least 80% less in response to the four types of LPS tested at wide dose ranges. Other inflammatory agents, such as proteose peptone, PHA, zymosan, or inulin failed to demonstrate differences between susceptible and resistant mice. In attempting to delineate the mechanism(s) responsible for the widely different responses to LPS, we found that in all strains the serum C titers were equal; the levels of chemotactic activity produced by LPS in sera in vitro were equal; and the chemotactic capabilities in vitro of the PMN were equal. In comparing the inflammatory responses generated in the resistant and susceptible mice to native endotoxin, lipid A-KDO glycolipid, and lipid A, it was determined that the carbohydrate moieties of the native endotoxin molecule were necessary for the preservation of the large differential inflammatory responses seen in the C3H/Hej and normal mice. Finally, we demonstrated that the survival of susceptible mice challenged with endotoxin i.p. could be enhanced by augmenting the magnitude of the i.p. inflammatory response via simultaneous injection of a second inflammatory agent. Thus, the rapid influx of inflammatory cells in the C3H/Hej mouse may be integrally related to its endotoxin resistance.
Replication of Herpes Simplex Virus and Cytomegalovirus in Human Leukocytes
Authors: Rinaldo Charles R. , Richter Bonnie S. , Black Paul H. , Callery Richard , Chess Leonard , Hirsch Martin S.
Published: December 2022

Human peripheral blood leukocytes, lymphocyte subpopulations, and hemic cell lines were examined for their ability to support HSV and CMV replication. Mitogen-stimulated mononuclear leukocytes, B lymphocytes, and T lymphocytes supported the replication of HSV to high titers over 3 to 5 days of infection. HSV replicated in unstimulated mononuclear leukocyte cultures of one of five donors, and to a limited degree in untreated B lymphocytes of three of five donors; HSV replication was not detected in unstimulated T lymphocytes (five donors). There was no evidence of enhanced uptake of 3H-thymidine in the untreated donor cells that replicated HSV. CMV replication was not detected during 9 to 10 days of infection in untreated or mitogentreated mononuclear leukocytes and lymphocyte subpopulations from the same adult donors or in neonatal cord blood leukocytes. The ability of the cells to support HSV or CMV replication did not correlate with the presence of specific antiviral antibodies in the donor serum.

HSV replicated in B, T, and myeloid cell lines to high titers over 5 days of infection, whereas CMV failed to replicate in any of the hemic cell lines. A persistent HSV infection has been established in a T cell line (CEM) with high titers of infectious virus being produced concurrently with growth of the cells over the first 11 weeks of infection.
Colonic Basophil Hypersensitivity
Authors: Askenase Philip W. , Boone Walter T. , Binder Henry J.
Published: December 2022

Contact hypersensitivity reactions were elicited by intraluminal application of DNCB to the colon of guinea pigs and were found sometimes to contain large infiltrates of basophils. These colonic basophil hypersensitivity reactions were accompanied by systemic sensitization and were not seen in nonspecific colonic inflammatory responses induced by croton oil. The heterogenous immune inflammatory responses induced in the colon by DNCB contact sensitivity may serve as a model for the study of inflammatory bowel disease.
Autochthonous, Allogeneic, and Xenogeneic Cells as Targets for Vaccinia Immune Lymphocyte Cytotoxicity
Authors: Woan Melissa C. , Yip Dick-Man , Tompkins Wayne A. F.
Published: December 2022

The cytotoxicity of vaccinia-immune rabbit spleen cells against autochthonous, allogeneic, and xenogeneic vaccinia-infected target cells was studied by using the 51Cr release assay. The results showed that whereas effector cells from rabbit spleen could not lyse the xenogeneic target cells, there was no consistent difference in lysis of autochthonous and allogeneic targets. Antibody-mediated cytotoxicity showed the resistance to cell-mediated lysis in this system not to be the result of reduced virus antigen on the cell surface. These data also showed the established cell line RK13 was markedly more sensitive to immune spleen cell lysis than short-term rabbit skin fibroblast cultures. The data suggested that it is possible to develop standardized tests to evaluate cell-mediated immunity to virus infections in a noninbred population.
Immunopathology of Experimental Allergic Orchitis in the Rabbit
Authors: Tung Kenneth S. K. , Woodroffe Andrew J.
Published: December 2022

Experimental allergic orchitis (EAO) was induced in the rabbit by immunization with a partially purified testicular antigen (TCA) from rabbit or guinea pig (GP) in complete Freund's adjuvant. All rabbits immunized with rabbit TCA (RTCA) developed severe testicular disease early (day 10) whereas serum antibody to rabbit sperm was detected late (day 60). Immune complexes, with one exception, were not found in the testicular lesions. In contrast, rabbits immunized once with GPTCA developed high titers of serum antisperm antibody early (day 10) in association with focal and mild testicular lesions that were inconsistent. However, when rabbits were hyperimmunized with GPTCA (three injections), severe orchitis developed in association with peritubular immune complexes. The immune complexes, composed of sperm antigen, antisperm antibody of IgG and IgM classes and C3, were confined to the testis. Antibody from rabbits immunized with RTCA reacted with rabbitspecific sperm antigens, whereas antibody from GPTCA-immunized rabbit reacted in part with sperm antigens that were shared between rabbit and GP. The results indicate that 1) EAO can be readily induced in the rabbit, 2) rabbit EAO can be induced with testicular antigens of homologous or heterologous origin, 3) GP and rabbit antigens elicit antisperm antibody with different specificities, 4) the immunopathology of EAO induced by rabbit and GP antigens differs and suggests involvement of different pathogenetic mechanisms, and 5) in GP antigen-induced EAO, the findings are indistinguishable from those of postvasectomy orchitis in the rabbit.
Antibody Response and Tumor Growth in Syngeneic Mice Immunized to Partially Purified B16 Melanoma-Associated Antigens
Authors: Bystryn Jean-Claude
Published: December 2022

Soluble murine melanoma-associated antigens (MAA), partially purified from the media of B16 melanoma cells in culture by ammonium sulfate precipitation and Sephadex G-200 chromatography, were tested for their effect on tumor growth. MAA were immunogenic in syngeneic mice as evidenced by their ability to induce anti-melanoma antibodies and partial protective tumor immunity. The level of immunity was variable. It ranged from retardation of tumor development to almost complete suppression of tumor growth. The results were influenced by the nature of the control group, since immunization to either normal tissue antigens or complete Freund's adjuvant enhanced tumor growth. Overall, 46 of 91 MAA immunized mice, but none of 114 control mice, survived over 6 weeks (p < 0.001). The protective immunity was specific since MAA immunized mice were not resistant to challenge with an unrelated syngeneic tumor (BW 10232 adenocarcinoma). Partially purified normal tissue antigens were also immunogenic in syngeneic mice. They induced low levels of antibodies to, and enhanced the growth of, melanoma.

These findings indicate that the culture media of melanoma cells contains tumor antigens that retain their biologic activity after partial purification, and that can induce specific, although only partial, protective immunity to melanoma.
Macrophage Ia Antigens
Authors: Cowing Carol , Schwartz Benjamin D. , Dickler Howard B.
Published: December 2022

By indirect immunofluorescence and microcytotoxicity it was demonstrated that different populations of murine macrophages bear different amounts of Ia antigens on their membranes. At least three subpopulations could be distinguished: those that lack Ia antigens and predominate in peritoneal exudate; cells bearing I-A antigens that are the majority of splenic macrophages and a minor population in the peritoneum; and cells bearing I-C antigens that are a minor population in both spleen and peritoneum. Internal radioisotope labeling studies confirmed that the I region molecules are synthesized by the macrophages. It is suggested that these different macrophage subpopulations may play distinct roles in the immune response.
Effects of Antigen-Feeding on Intestinal and Systemic Immune Responses
Authors: Kagnoff Martin F.
Published: December 2022

The ability of antigens administered by the intestinal route to alter the cytotoxic T lymphocyte (CTL) population in intestinal and extraintestinal sites was examined by feeding mice tumor cells bearing H-2 or non-H-2 genecoded cell surface alloantigenic differences. Peyer's patches from mice fed tumor cells differing at H-2 demonstrated a 12-fold increase in specific CTL activity over controls after stimulation in culture but the precursor CTL population in Peyer's patches was not expanded by feeding mice cells bearing minor H gene coded alloantigenic differences. In contrast, the precursor CTL pool in spleen was increased both in mice fed cells bearing alloantigenic differences coded for by H-2 and non-H-2 gene loci. The precursor CTL population in mesenteric lymph nodes (MN) was not expanded significantly by tumor cell feeding. Feeding was nearly as effective as i.p. priming at expanding the Peyer's patch precursor CTL population when tumor cells bearing H-2 differences were used, whereas i.p. priming was more effective than feeding in expanding the precursor CTL population in spleen.
Dual Binding Specificities in Mopc 384 and 870 Murine Myeloma Immunoglobulins
Authors: Bhattacharjee Apurba K. , Glaudemans Cornelis P. J.
Published: December 2022

Homogeneous murine myeloma immunoglobulins (IgA, κ), M 384, and M 870, bind methyl α-D-galactopyranoside and phosphorylcholine at different subsites. Heterologous recombinant immunoglobulins of these two immunoglobulins with M 603 (a homogeneous IgA, κ with known phosphorylcholine specificity) also bind phosphorylcholine.
The Genetic Mapping of a Defective Lps Response Gene in C3H/HeJ Mice
Authors: Watson James , Kelly Kathleen , Largen Michael , Taylor Benjamin A.
Published: December 2022

The expression of a defective LPS response gene Lps and the major urinary protein (Mup-1) are concordantly inherited in backcross (C3H/HeJ × C57BL/6J)F1 × C3H/HeJ mice, indicating genetic linkage of these loci. Mup-1 is known to be linked to the brown coat color locus on chromosome 4 in mice; thus Lps can now be assigned to chromosome 4. A value of 0.06 ± 0.02 has been estimated for the recombination frequency between Mup-1 and Lps. We have used the polysyndactyly (Ps) mutation further to localize Lps on chromosome 4. Lps is located between the Mup-1 and Ps loci.
Generation of T Helper Cells in Vitro
Authors: McDougal J. Steven , Cort Sheila P.
Published: December 2022

A sequential culture technique for the in vitro induction and subsequent assay of T helper cells is employed to examine the histocompatibility requirements for antigen recognition by murine T helper cells. F1 T cells are primed in vitro with antigen-pulsed parental strain macrophages and tested for antigen-specific helper activity in cultures containing anti-Thy 1.2 serum and C treated spleen cells from hapten-primed parental or F1 mice. A semiallogeneic system is used and appropriate controls are included to avoid possible complicating effects of allogeneic interactions. The results indicate that F1 T helper cells preferentially stimulate carrier-specific anti-hapten plaque-forming cell responses in spleen cells which are H-2 identical with the macrophage used initially to prime the T cells. Parental spleen cell cultures do not respond to F1 T helper cells which were primed with the other parental strain macrophage. Supplementing this culture with macrophages which are histocompatible with those used to prime the F1 T cells is sufficient to restore T helper cell activity. Thus, the genetic restriction described here is between the primed T cell and the macrophage used to elicit secondary responses and not between the T cell and B cell. The results in this semiallogeneic system, however, do not rule out the possibility of additional allogeneic genetic restrictions in the subsequent interaction of T cells with B cells.
Lactic Dehydrogenase Virus Alteration of Lymphocyte Circulation
Authors: Mongini Patricia K. A. , Rosenberg Leon T.
Published: December 2022

Acute infection with lactic dehydrogenase virus (LDV) causes a systemic alteration in lymphocyte circulatory patterns. Peripheral lymph nodes (LN) and spleens in acutely, but not chronically, infected mice retain a significantly greater proportion of injected 51Cr-labeled lymphocytes than the respective tissues in noninfected controls. This increase in lymphocyte localization in LN and spleen is dependent upon the dose of LDV injected and the timing of the infection. A relatively large dose of LDV (108 infectious units) causes an early but very transient increase in splenic lymphocyte localization accompanied by an early but prolonged increase in lymphocyte recovery in LN. Smaller doses of LDV cause more prolonged effects on splenic lymphocyte recovery and retarded effects on lymphocyte localization in LN. Increases in splenic recovery were always accompanied by decreases in hepatic recovery of lymphocytes. LDV-induced alteration in lymphocyte circulation may be responsible for many previously observed modifications of immune responses in LDV-infected mice.
Mode of Action of a Soluble Immune Response Suppressor (Sirs) Produced by Concanavalin A-Activated Spleen Cells
Authors: Tadakuma Takushi , Pierce Carl W.
Published: December 2022

Soluble immune response suppressor (SIRS) is a product of concanavalin A activated Ly 2,3+ T cells which suppresses plaque-forming cell response to sheep erythrocytes by murine spleen cells in vitro through its action on macrophages. The mechanism(s) by which SIRS-treated macrophages suppress plaque-forming cell responses has been investigated. Experiments with varying ratios of control supernatant- and SIRS-treated macrophages showed that suppression was not due to inactivation of macrophages, but was an active process. This active process required at least 3 days for full expression since: 1) responses in cultures containing SIRS-treated macrophages developed normally for 3 days and suppression became apparent only on day 4, approximately 24 hr after a significant decrease in DNA synthesis; 2) responses of spleen cells in a Millipore chamber which were incubated with SIRS-treated macrophages for 48 hr and then transferred to control supernatant-treated macrophages for the last 72 hr of culture escaped suppression; and 3) addition of SIRS-treated macrophages to spleen cell cultures after initiation resulted in later expression of suppression. The experiments in which spleen cells were incubated in Millipore chambers separated from SIRS-treated macrophages indicated that a second factor(s), released from the SIRS-treated macrophages, was active in suppressing the response. This factor may also be responsible for the decrease in DNA synthesis after day 3 of culture and suggests that SIRS-mediated suppression is ultimately due to limitation of proliferation of responding B cells.
Agents That Increase Cyclic AMP Inhibit Accumulation of cGMP and Depress Human Monocyte Locomotion
Authors: Gallin John I. , Sandler Jeffrey A. , Clyman Ronald I. , Manganiello Vincent C. , Vaughan Martha
Published: December 2022

Ascorbic acid and serotonin increased the 3′,5′-guanosine monophosphate (cGMP) content and enhanced the chemotactic responsiveness of mononuclear cells from human peripheral blood. The ionophore A23187, PGE1, and polystyrene beads increased the 3′,5′-adenosine monophosphate (cAMP) content of mononuclear cells and had no effect on basal cGMP content. A23187, PGE1, and beads caused significant increases in cAMP content in preparations of adherent cells (chiefly monocytes); PGE1 and beads also increased cAMP content in the nonadherent cells (chiefly lymphocytes). A23187 and PGE1 each inhibited mononuclear cell locomotion. The chemically unrelated agents, A23187, beads, and PGE1, that raised cAMP of monocytes inhibited the accumulation of cGMP in response to serotonin and asorbic acid. PGE1 at a concentration that caused a 349% increase in cAMP inhibited the effect of serotonin on cGMP accumulation and the amplification of chemotaxis by serotonin only 37% and 40%, respectively. In contrast A23187, which increased monocyte cAMP by only 71%, inhibited the effect of serotonin on cGMP accumulation and locomotion by 85 and 79%, respectively. Although there is no clear quantitative relationship between the increase in cAMP content and the degree of inhibition of cGMP accumulation, it is possible that the elevation of cAMP in monocytes interferes with their capacity to accumulate cGMP in response to serotonin and ascorbic acid, perhaps by increasing cGMP degradation and/or inhibiting synthesis. In any case cyclic nucleotide modulation of monocyte locomotion appears to be more closely related to intracellular cGMP content than to cAMP content.
Human Cell Membrane Components Bound to β2-Microglobulin in T Cell-Type Cell Lines
Authors: Tada Nobuhiko , Tanigaki Nobuyuki , Pressman David
Published: December 2022

Cell membrane components bound to β2-microglobulin were isolated from Renex 30 (a nonionic detergent)-solubilized membrane materials of two human T cell-type cell lines, MOLT-4 and CCRF-CEM, by gel filtration and lectin affinity chromatography. The isolation was carried out by following the β2-microglobulin activity by radioimmune inhibition assay. The T cell membrane components bound to β2-microglobulin had a uniform molecular size of about 200,000 daltons and most of them showed an affinity to lentil lectin. The isolated membrane components were radioiodinated and examined for identity to HLA antigens by sequential precipitation with rabbit anti-HLA antiserum (specific to HLA large components) and with rabbit anti-β2-microglobulin antiserum. In addition to HLA antigens, the β2-microglobulin-bound components obtained from the MOLT-4 cells were found to contain certain membrane components that are the same in molecular size as the HLA large components but that are different antigenically from the HLA large components. On the other hand, the β2-microglobulin-bound membrane components obtained from the CCRF-CEM cells were all HLA antigens. No other membrane components were involved in the binding.
Specific Functional and Immunologic Assay of Plasma Plasminogen in Hereditary Angioedema, in Hereditary Angioedema Treated with Tranexamic Acid, and in Normal Subjects
Authors: Spragg Jocelyn
Published: December 2022

Plasma plasminogen levels were determined by a specific esterolytic assay and a radial immunodiffusion assay, both of which were standardized on a weight basis by using highly purified plasminogen diluted in plasminogen-free plasma. In thirteen normal individuals the functional and antigenic levels were 275 ± 61 µg/ml and 285 ± 61 µg/ml, respectively (r = 0.95), whereas in 58 plasmas from individuals with hereditary angioedema the levels were 272 ± 58 µg/ml and 265 ± 64 µg/ml, respectively (r = 0.89). In two patients treated with tranexamic acid either acutely or chronically, both the functional and antigenic plasminogen levels were diminished.
Myeloma-Specific Transplantation Resistance: a Requirement for Complete Freund's Adjuvant Stimulation of Effectors
Authors: Bridges Sandra H.
Published: December 2022

BALB/c mice immunized with purified BALB/c myeloma protein M315 produce anti-idiotypic antibody (a-Id315), directed predominantly to the binding site of that protein, and are resistant to challenge with the corresponding plasmacytoma, MOPC-315. Previous studies showed that the induction thresholds and decay patterns for the two activities were very similar; in the present study, the two activities were separated. Mice that received their initial exposure to M315 in incomplete Freund's adjuvant (IFA), instead of complete Freund's adjuvant (CFA), produced a-Id315 but transplantation resistance was greatly reduced in magnitude. With respect to IgG subclass distribution and hapten inhibitability by ε-DNP-L-lysine, the specific ligand of M315, the antibody was indistinguishable from that produced when CFA was used as adjuvant. Although the binding capacity of the antibody was in general lower than that produced with the standard protocol with CFA, examination of the binding capacities of sera from individual IFA-immunized mice demonstrated that the diminished resistance was not strictly correlated with the level of binding capacity. When a-Id315 activity was monitored in the sera of M315-immunized mice after tumor cell challenge, it was found that IFA-immunized mice maintained antibody levels as well as CFA-immunized mice. In mice developing tumors during the monitoring period, activity was maintained until 2 to 4 days before the detection of palpable tumors or M315 protein in the serum. The results suggest that although a-Id315 may be involved in the mediation of the myeloma-specific transplantation resistance, other elements of the immune system, which are stimulated by CFA, are required for the full expression of resistance.
Nonspecific Inhibition of Tumor Growth by Allosensitized Lymphocytes
Authors: Nakayama M. , Sendo F. , Miyake T. , Fuyama S. , Arai S. , Kobayashi H.
Published: December 2022

In a previous paper we demonstrated with the Winn test that rat spleen cells sensitized to an allogeneic tumor (AH-66 from Donryu strain) nonspecifically inhibited in vivo growth of a syngeneic tumor (KMT-17 from WKA strain) only when the spleen cells were stimulated with inactivated AH-66 cells used for immunization. These results were obtained with spleen cells harvested 2 weeks after immunization.

In the present study with the Winn test, when tumor inhibitory activity of AH-66-sensitized spleen cells was serially examined after immunization, the early spleen cells (1 week after immunization) inhibited growth of KMT-17 without further addition of inactivated AH-66 cells, but the late spleen cells (2 weeks after immunization) did not. AH-66-sensitized spleen cells 60 days after immunization did not inhibit growth of KMT-17 even with further addition of inactivated AH-66 cells, whereas these spleen cells inhibited in vivo growth of homologous AH-66. On the other hand, the early spleen cells alone did not inhibit growth of admixed KMT-17 when the mixtures were in vivo cultured in diffusion chambers where no inflammatory cells existed. However, further addition of polymorphonuclear leukocytes (PMN) to the early spleen cells reduced the number of viable KMT-17 cells even in diffusion chambers. The late spleen cells did not inhibit KMT-17 growth in diffusion chambers even when PMN were further added. Normal thymocytes, spleen cells or peritoneal macrophages did not cooperate with the early spleen cells in inhibiting the growth of KMT-17.
Expression of Group b Light Chain Allotypes in Cottontail Rabbits
Authors: Smith Linda J. , Mandy William J.
Published: December 2022

Group b allotypic determinants (b4, b5, b6, and b9) were detected on cottontail IgG by several assays including immunodiffusion, radioimmune binding, and inhibition of radiobinding. The results indicate that cottontail IgG possess some but not all of the subspecificities present on domestic rabbit IgG. Several cottontail rabbits exhibited three of the four possible allotypic markers. In these instances, however, only two populations of IgG molecules (i.e., phenogroups) could be detected, each bearing one, two, or three of the allotypes. Five separate and distinctive phenogroups were identified. The results suggest that the phenogroups represent products of multiple allelic genes for the constant region of cottontail rabbit κ light chain.
Cell-Mediated Lympholysis of Trinitrophenyl-Derivatized Autologous Human Cells: in Vitro Triggering by Nonspecific Signals
Authors: Friedman Steven M. , Neyhard Nancy , Chess Leonard
Published: December 2022

This report describes the primary in vitro generation of human CTL that lyse TNP-derivatized autologous cells. Although in the majority of these studies, a direct cytotoxic response to the TNP-modified autologous stimulators was not achieved, in all experiments the addition of either allogeneic cells or soluble antigen triggered the generation of killer cells which destroy TNP-modified, but not unaltered, autologous targets. Fractionation of responder lymphocyte populations demonstrated that the cytotoxic activity was mediated by T cells. Killer cell specificity was tested by assaying for cytotoxicity to a variety of targets, and by blocking the cytolysis of TNP-altered autologous targets with various populations of nonradiolabeled cells. Results indicated that these CTL were cytotoxic for TNP-modified autologous cells but not unaltered autologous or TNP-modified allogeneic targets. The capacity of soluble antigen and alloantigens to facilitate the in vitro generation of altered-self reactive human CTL is not an isolated phenomenon. This “helper” effect has now been observed for the cytotoxic response to chemically modified autologous cells and MHC identical human leukemic blasts. It is possible that in vivo, similar responses to nonspecific antigenic stimuli may play a role in the maintenance of immune surveillance.
Nature of the Antigenic Complex Recognized by T Lymphocytes
Authors: Thomas David W. , Shevach Ethan M.
Published: December 2022

Nonimmune (2 × 13)F1 guinea pig T lymphocytes initially stimulated with trinitrophenyl- (TNP) modified macrophages from one parental strain and then treated with bromodeoxyuridrine (BUdR) and light are unable to be primed subsequently with TNP-modified macrophages of the same parental strain. In contrast, these BUdR and light treated F1 T cells can be primed with TNP-modified macrophages of the other parental strain. These results demonstrate that in a nonimmune F1 animal two T cell subpopulations exist before priming that are genetically predisposed to respond to antigen associated with macrophages derived from one or the other parent.
Characterization of a Fetal Calf Serum-Derived Molecule Reactive with Human Natural Antibodies: Its Occurrence in Tissue Culture-Grown Type C RNA Viruses
Authors: Snyder Harry W. , Fox Michelle
Published: December 2022

By means of a sensitive radioimmunoprecipitation (RIP) assay, simian sarcoma virus-simian sarcoma-associated virus (SSV-SSAV), purified from culture fluids of infected normal rat kidney (NRK) cells, was shown to acquire a surface antigen from serum used in the tissue culture medium. This antigen, which was acquired when serum from either fetal calf, horse, swine, rabbit, or chicken origin was used, accounted for a substantial portion (but not all) of the total precipitating activity exhibited by natural human antibodies for membrane-associated antigens of these viruses. By 1) alcohol precipitation, concanavalin A chromatography, and Sephadex G-150 filtration of fetal calf serum (FCS) proteins or 2) chromatography of serum proteins over a human IgG-containing immunoaffinity column, a glycoprotein of approximately 55,000 daltons has been identified which is a minor constituent of FCS (<0.1% of total protein) and has the antigenic capacity of whole FCS.
Dissociated and Reconstituted Subcellular Alloantigen Capable of Stimulating Mouse Cytotoxic T Lymphocytes in Vitro
Authors: Fast Loren D. , Fan David P.
Published: December 2022

Reconstituted subcellular membrane fragments were prepared from sonicated murine lymphoid tissue membrane fragments by detergent solubilization and dialysis. The reconstituted subcellular antigen was compared with sonicated cell membranes and x-irradiated cells for the ability to stimulate the formation of allogeneic cytotoxic thymus-dependent lymphocytes in vitro. The results showed that the reconstituted subcellular antigen had properties similar to those reported for the sonicated antigen. Both types of antigen could stimulate immune spleen cells but not normal spleen cells. Furthermore, soluble H-2 antigens, even of membrane origin, were incapable of the stimulation unless reconstituted into membrane fragments large enough to be centrifuged at 48,000 × G for 30 min.
The Induction of Cytolytic T Lymphocytes with Purified Plasma Membranes
Authors: Lemonnier François , Mescher Matthew , Sherman Linda , Burakoff Steven
Published: December 2022

Subcellular fractions were demonstrated to be effective in stimulating primary and secondary allogeneic CTL responses. 5′ nucleotidase activity was used to assess purification of plasma membranes and stimulating activity was found to co-purify with the plasma membrane fraction of the cell. Electron micrographs of these purified plasma membranes showed the majority of the material to be in the form of vesicles of relatively heterogeneous size. The cytolytic T lymphocytes generated by incubation with purified plasma membranes demonstrated immunologic specificity.
Role of T Lymphocytes in Adjuvant Arthritis
Authors: Kayashima Kohji , Koga Toshitaka , Onoue Kaoru
Published: December 2022

The effects of treatments with cyclophosphamide (CY), hydrocortisone and anti-thymocyte sera (ATS) on the development of adjuvant arthritis (AA) were examined in WKA rats inoculated with wax D to induce AA. A single injection of 25 to 50 mg/kg of CY given 2 to 3 days before wax D inoculation caused severe arthritis with high incidence, whereas larger doses of CY were less efficient. Furthermore, the enhancing effect of CY pretreatment was abolished by passively transferred normal syngeneic thymocytes 1 day before wax D inoculation, but not by thymocytes that had been treated with ATS and guinea pig C. On the basis of these results and the previous observations on the effect of adult thymectomy and low-dose irradiation, we concluded that this enhancing effect of CY pretreatment was caused by selective depletion of suppressor T lymphocytes. Pretreatment with 12.5 mg of hydrocortisone also caused severe arthritis. This enhancing effect of hydrocortisone could also be due to the elimination of those suppressor cells. In contrast, in vivo pretreatment with ATS showed striking inhibition on the development of AA, suggesting that ATS could eliminate T lymphocytes that were responsible for eliciting this disease. Thus it appears that at least two T cell subpopulations are involved in the development of AA, one is an ATS-sensitive T2 subpopulation that is effective for induction of AA and the other is a T1 subpopulation that regulates this disease process.
The Analysis of the Monoclonal Immune Response to Influenza Virus
Authors: Gerhard Walter
Published: December 2022

Individual splenic precursor B cells from BALB/c mice primed with influenza virus PR8[A/PR/8/34 (H0N1)] were stimulated in vitro in the splenic fragment culture system by homologous or various heterologous influenza viruses. The specificity of the stimulated precursor cells was determined by analysis of the antibodies secreted by the ensuing plasma cell clone in a radioimmunoassay (RIA).

Viruses of the H2N2 and H3N2 subtypes were unable to stimulate hemagglutinin (HA)- or neuraminidase (NA)-committed precursor B cells but did efficiently stimulate chicken host component (ChHC)-committed precursors. Viruses of the H1N1 and H0N1 subtypes could stimulate precursors committed to any of the three viral surface components. Analysis of the fine specificity of HA-committed B cells showed that BEL(H0N1) and CAM(H1N1) stimulated almost exclusively precursors whose clonal antibody product reacted with the stimulating virus in the RIA. On the other hand, WSE and MEL (both H0N1) quite frequently were able to stimulate precursors whose clonal antibody product did not react with the stimulating virus in the RIA. These results suggest that the stimulatory interaction of viruses with the cell-bound immunoglobulin receptors is slightly less affinity dependent than the antigen-antibody interaction in the RIA.
Generation of Suppressor Lymphocytes During Sensitization in Culture against a Syngeneic Tumor: Affinity Chromatography on Insolubilized Histamine
Authors: Schechter Bilha , Segal Shraga , Feldman Michael
Published: December 2022

We investigated the effect of depletion of histamine-binding lymphoid cells on immunological properties of lymphocytes sensitized in culture against tumor cells. C57BL/6 spleen cells that were sensitized in vitro on monolayers of the syngeneic Lewis lung carcinoma (3LL) became cytotoxic to the tumor cells in vitro after 3 to 5 days of sensitization. Sensitized cells harvested after 4 days of sensitization occasionally enhanced tumor growth in vivo. Fractionation of the sensitized lymphocytes over insolubilized histamine-rabbit serum albumin-Sepharose (HRS) columns decreased or abolished the enhancing activity in vivo and specifically increased the in vitro cytotoxic activity of the depleted lymphocytes. A similar increase in the cytotoxic activity of HRS-fractionated cells was observed in an allogeneic combination of C57BL spleen cells sensitized against C3H fibroblasts. The effect of HRS chromatography on the in vitro cytotoxic activity increased with prolonged incubation of the depleted effector cells with the target cells.
Suppression of the Mitogen-Stimulated Blastogenic Response during Reticuloendotheliosis Virus-Induced Tumorigenesis: Investigations into the Mechanism of Action of the Suppressor
Authors: Carpenter Charles R. , Rubin Arnold S. , Bose Henry R.
Published: December 2022

Splenic lymphocytes from chickens infected with reticuloendotheliosis virus (REV) are suppressed in their ability to undergo PHA-induced blastogenesis. This suppression can be detected within 72 hr after virus injection and requires active viral infection since i.p. or i.v. injection of UV-inactivated REV does not result in inhibition of the blastogenic response. Suppressor cells from the spleens of REV-infected birds severely inhibit the ability of spleen cells from uninfected chickens to respond to PHA at a ratio of 1:20, suggesting that each suppressor cell may be capable of suppressing more than one target cell. Contact between suppressor and target cells is required for the rapid inhibition of the normal PHA response. The suppressive mechanism is cytostatic in nature, and apparently of host origin since neither REV, nor REV-infected (or transformed) cells mediate the suppression. The ability of the suppressor cells to impair the blastogenic response of spleen cells is sensitive to trypsin, suggesting that an inhibitory protein is exposed on the surface of the suppressor cells.
Inhibition of in Vivo and in Vitro Neutrophil Responses to Chemotactic Factors by a Competitive Antagonist
Authors: O'Flaherty Joseph T. , Showell Henry J. , Kreutzer Don L. , Ward Peter A. , Becker Elmer L.
Published: December 2022

Carbobenzoxy-phenylalanyl-methionine (CBZ-Phe-Met) is an inhibitor (antagonist) of the binding to the receptor on polymorphonuclear neutrophils (PMN) for a series of synthetic chemotactic oligopeptides. This inhibitor was studied for its effects on the following chemotactic factor-induced PMN functions: a) chemotaxis, b) granule enzyme release, c) cellular aggregation, and d) in vivo induction of neutropenia. Each of these functions when induced by the chemotactic tripeptides (formyl-methionyl-leucyl-phenylalanine or formyl-methionyl-methionyl-methionine) was inhibited by the antagonist. For each in vitro assay the concentration of antagonist causing a 50% inhibition of response was approximately 10-4 M and the dissociation constant for the antagonist was approximately 10-5 M. Inhibition was dose related, reversible, and competitive. In contrast to these results, the antagonist, CBZ-Phe-Met, did not inhibit any of the four PMN responses induced by the chemotactic fragment of C5. These results further strengthen the relationship between all four chemotactic factor-induced PMN responses and suggest that these chemotactic tripeptides induce each response by interacting with a single receptor. Antagonists of chemotactic factors may represent a new approach to the in vivo suppression of leukotactic factor-mediated biologic responses.
Two Functionally Distinct Subpopulations of Human T Cells That Collaborate in the Generation of Cytotoxic Cells Responsible for Cell-Mediated Lympholysis
Authors: Evans Robert L. , Lazarus Herbert , Penta Ann C. , Schlossman Stuart F.
Published: December 2022

A human thymus-dependent differentiation antigen, TH2 was defined by a rabbit anti-human T cell serum absorbed with autologous B lymphoblasts and leukemic cells bearing T cell markers from a patient with chronic lymphocytic leukemia. Anti-TH2 reacted specifically with thymus-derived lymphoid cells and exhibited two distinct profiles of reactivity with normal peripheral T cells as detected by indirect immunofluorescence on a FACS I. Isolation of strongly reactive, TH2+, from weakly reactive, TH2- T cells by fluorescence-activated cell sorting revealed that the TH2+ subset contained most of the killer activity in cell-mediated lympholysis (CML), but had a diminished response in MLC and a suboptimal or negligible proliferative response to soluble antigens (mumps, PPD, tetanus toxoid). In contrast, the TH2- subset contained markedly less killer activity but amplified cytotoxicity by TH2+ cells and exhibited a proliferative response to both alloantigen and soluble antigens that was often significantly greater than the response by unseparated T cells. The relevance of these findings to previously described human T cell subsets and to functional subpopulations of murine T cells is discussed.
Suppression of the Generation of Human Con A-Induced Cytotoxic Effector Cells by Con A-Activated Suppressor Cells
Authors: Hunninghake Gary W. , Fauci Anthony S.
Published: December 2022

The present study demonstrates that in vitro stimulation by Con A of peripheral blood lymphocytes from certain normal individuals may result in preferential activation of either cytotoxic effector cells capable of killing allogeneic target cells or activation of suppressor cells that prevent the generation of cells cytotoxic to allogeneic targets. Con A-activated lymphoid cells from individuals displaying high levels of cytotoxicity against allogeneic targets (high responders) did not suppress the generation of cytotoxic cells in cell suspensions from other high responders; however, Con A-activated cells from individuals displaying low levels of killing against allogeneic targets (low responders) regularly suppressed the generation of cytotoxic cells in cell suspensions from high responders. Lymphoid cells from low responders suppressed the generation of cytotoxic cells when cocultured in the presence of Con A with cells from a high responder but did not affect the function of killer cells once they had been generated. Pretreatment of cells from a low responder with mitomycin C before co-culturing them in the presence of Con A with lymphoid cells from a high responder impaired their ability to suppress the generation of cytotoxic cells, suggesting that proliferation was necessary for optimal generation of these suppressor cells. In addition, it was demonstrated that Con A-activated cells from low responders were unable to kill an allogeneic target, even when these cells were generated in the presence of a high responder, suggesting that a lack of helper cells was not a sufficient explanation for the inability of cells from low responders to generate killer cells. These studies provide further evidence that suppressor cell functions of subpopulation of lymphoid cells play an important role in the regulation of immune responses of normal individuals.
Linkage between a Gene for a Serum Protein and the Gene for the Ig Kappa Light Chain in Rabbits
Authors: Usher David C. , Mayer Eugene P. , Dray Sheldon , Reiter Harvard
Published: December 2022

A rabbit serum protein (Prt) that migrates ahead of transferrin in polyacrylamide gel electrophoresis is inherited in a simple Mendelian fashion. The gene for this protein is linked to the gene for the Ig κ light chain and the degree of linkage is calculated as 23.5 ± 4.7 centimorgans.
Partial Purification and Molecular Characterization of a Lymphokine (Costimulator) Required for the Mitogenic Response of Mouse Thymocytes in Vitro
Authors: Shaw Jennifer , Monticone Vicky , Paetkau Verner
Published: December 2022

We have partially purified a lymphokine, costimulator, which is necessary to induce mitogenesis in mouse thymocytes in vitro. Costimulator is released from mouse leukocytes exposed to Con A for 12 to 18 hr. It has been purified more than 100 X by gel exclusion chromatography and isoelectric focusing. Thymocytes from CBA/J mice respond to the mitogenic lectin Con A only if the costimulator concentration is above a certain level. Culturing such cells with Con A at a density below 1 × 106 cells/ml produces costimulator concentrations too low for mitogenesis. This system has been developed into a quantitative assay for costimulator, to monitor purification, recovery, and biologic activity in various methods of molecular characterization. The activity is trypsin sensitive, and has a buoyant density characteristic of protein or glycoprotein. However, for a protein, it is relatively heat stable. Its m.w., established by carrying out sedimentation, gel filtration, and buoyant density measurements, is 30,500, and its frictional coefficient is 1.45. Costimulator purified by isoelectric focusing is active at 10-10 M or lower in tissue culture.
The Response to Mitogens and Allogeneic Cells of Human Lymphocyte Subpopulations Separated by Their Differential Binding to Monolayers of Bacteria
Authors: Kleinman Raphael , Teodorescu Marius
Published: December 2022

Various strains of bacteria were previously shown to bind to different lymphocyte subpopulations, i.e., to B cells and to four T cell subpopulations (T1, T2, T3, T4, our classification). We prepared bacterial monolayers and used them to separate lymphocyte subpopulations according to their binding properties. Bacteria were coupled to glutaraldehyde-treated gelatin layers and were subsequently fixed with formaldehyde. Suspensions of peripheral blood lymphocytes (PBL) were centrifuged against the bacterial monolayers and the cells that did not attach were resuspended with the help of a rocking platform. The percentage of PBL that remained attached was similar to the expected values according to the percentage of PBL-binding bacteria in a “rosette formation” assay. The cells that did not adhere to Escherichia coli-0 monolayers, i.e., practically all PBL, responded to the same extent as the unseparated PBL to four mitogens: phytohemagglutinin, concanavalin A, pokeweed mitogen, and streptolysin 0-associated mitogen. The lymphocytes that did not adhere to E. coli-2 monolayers, namely the T3T4 cells, responded poorly to all four mitogens. Also, the response to various doses of concanavalin A of T3T4 cells was much lower than that of purified T cells. The poor response of T3T4 cells was not due to the reduction in viability or to monocyte depletion. Moreover, the T3T4 cells responded in one-way mixed lymphocyte reactions as well as the unseparated PBL. We concluded that T3T4 lymphocytes could be separated by adherence to bacterial monolayers and that they may be functionally different from the other lymphocyte subpopulations identified by binding of bacteria.