bluuebunny/crossref_metadata_2025 huggingface dataset:
In Vitro Killing of Schistosomula of Schistosoma Mansoni by BCG and C. Parvum-Activated Macrophages
Authors: Mahmoud Adel A. F. , Peters Pierre A. S. , Civil Richard H. , Remington Jack S.
Published: December 2022

Resistance to Schistosoma mansoni infection in the mouse has been induced either specifically by a primary infection with this parasite or nonspecifically by a variety of immunostimulants such as BCG. In the present study we developed an in vitro system to examine the effector mechanism of nonspecifically induced resistance. Activated macrophage monolayers obtained from BCG- or Corynebacterium parvum-treated mice killed a respective mean of 32 ± 6% and 48 ± 5% of schistosomula after 24 hr incubation. The killing of the parasites was verified by their inability to mature to adult worms upon injection into normal mice. The activated macrophage-mediated killing was related to cell:parasite ratio, and was partially lost if the macrophage monolayers were kept in cultures for 24 hr before incubation with the organism. Supernatants of macrophages cultured in the presence of schistosomula killed a mean of 51 ± 3% of the organisms whereas those from cells cultured alone resulted in a mean killing of 25 ± 3%. Furthermore, toxic supernatants could be generated equally well on incubation with S. mansoni schistosomula or Trichinella spiralis larvae. Our data show that activated macrophage monolayers through soluble mediators destroy a significant proportion of the multicellular parasite S. mansoni schistosomula in vitro.
An Allotypic Marker on Chicken Immunoglobulin Light Chains
Authors: Foppoli James M. , Benedict Albert A.
Published: December 2022

The first chicken immunoglobulin light (L) chain allotypic specificity (L-1.1) to be described that was present on IgM, 7S Ig, Fab, and L chains was detected by radioimmunoassay. The gene controlling the expression of L-1.1 is inherited in a simple Mendelian fashion at an autosomal locus and is unlinked to a constant region heavy chain locus, four blood group loci and three loci determining lymphocyte cell surface alloantigens.
Macrophage Cell Lines Produce a Cytotoxin
Authors: Aksamit Robert R. , Kim K. Jin
Published: December 2022

Cytotoxic activity, measured by release of tritium from 3H-(methyl)-thymidine-labeled 7943 cells, was found in culture fluids from four of five macrophage cell lines. In the P388D1 cell line, cytotoxic activity was highest after 7 days of culture, 2 days after the cells reached confluence. The activity was not lost upon dialysis or lyophilization. When culture fluids were chromatographed on Sephadex G-200, two activity peaks were obtained: one in the void volume and the other corresponding to a m.w. of 40,000 to 50,000.
The Effects of Neuraminidase and Galactose Oxidase on Murine Lymphocytes
Authors: Kuppers Rudolf C. , Henney Christopher S.
Published: December 2022

Normal murine spleen cell populations treated sequentially with neuraminidase (NA) followed by galactose oxidase (GO) failed to mount cytotoxic responses to alloantigen. Such unresponsiveness was shown to be due to the induction of a suppressor T cell after NAGO treatment. Suppressor cell induction did not require cell proliferation, although mitogenic concentrations of galactose oxidase (0.019 units/ml after 50 units/ml NA) were needed. The NAGO-induced suppressor T cell was shown to be capable of suppressing both primary and secondary cytotoxic responses to alloantigen and to develop more readily in young (< 4-month-old) C57BL/6 spleen cell populations than in spleens from older mice. The suppressive activity of NAGO-treated spleen cells increased when they were cultured (1 to 3 days) before testing.

In contrast to the antigen-unresponsiveness of normal spleen cells after NAGO treatment, spleens from alloantigen-primed mice still responded vigorously to the priming antigen after NAGO treatment, although their primary responses to other alloantigens were suppressed. Antigen priming per se did not markedly affect the induction of suppressor T cells by NAGO; the differences between the ability of normal vs primed spleen cells to mount cytotoxic responses after NAGO treatment seemed largely to reflect the relative insusceptibility of secondary cytotoxic responses to the action of suppressor T cells.
Lung Tumor-Reactive Cytotoxic Lymphocytes Generated in Mixed Lymphocyte Reaction Between C3HfeB/HeN (H-2kb) and C3H/HeN (H-2k) Strain Mice
Authors: Imamura Masahiro , Gipson Thomas G. , Bensky Norman , Justice Robert , Martin W. John
Published: December 2022

The tumor-associated transplantation antigen expressed by several transplacentally induced lung tumors of C3HfeB/HeN mice (H-2kb haplotype) has previously been shown to exist as a normal tissue alloantigen in mice of known H-2k and H-2α haplotypes. This antigen is not expressed in normal tissues of C3HfeB/HeN mice but is expressed in C3H/HeN mice, the strain from which the C3HfeB/HeN mice were originally derived. The present study indicates that spleen cells from C3HfeB/HeN and C3H/HeN mice respond reciprocally in the mixed lymphocyte reaction. Cytotoxic T lymphocytes specific for the tumor-associated alloantigen can be readily generated in mixed lymphocyte reactions in which spleen cells from C3HfeB/HeN mice are reacted with x-irradiated spleen cells from C3H/HeN or A strain mice. These cells are effective in suppressing the growth of the C3HfeB/HeN-derived lung tumor 85 in x-irradiated syngeneic recipients.
Measurement of Human Type-Specific Antibody to Adenovirus Type 5 Hexon and Fiber Using a Fab′-Blocked Antigen Probe
Authors: Scott Jane V. , Dreesman Gordon R. , Kasel Julius A.
Published: December 2022

A radioimmunoassay (RIA) has been developed whereby antibodies to the type-specific antigenic determinants on the hexon or fiber subunits of adenovirus type 5 (Ad5) are selectively measured in the presence of group- and subgroup-reactive antibodies. To achieve this purpose, Fab′ fragments from rabbit antibody prepared against the most closely related serotype, Ad6, were used to block group- and subgroup-reactive determinants on the Ad5 proteins. The Ad6 Fab′ fragments were preincubated with purified iodinated Ad5 fiber or hexon. These Fab′-blocked 125I-labeled subunits were then used as type-specific antigenic probes in a conventional RIA. Serum specimens obtained from 20 volunteers, obtained before and after vaccination with purified Ad5 hexon or fiber subunit vaccine, were tested for antibody by using the Fab′-blocked Ad5 antigen probes. The results obtained demonstrated that i) the two noncross-reacting type-specific antibodies made against fiber or hexon are selectively detected by this procedure, ii) the relative mean concentrations of type-specific antibody ranged from 0.31 to 0.41% and 4.0 to 4.1% of the total antibody for hexon and fiber antigen, respectively, and iii) the levels of antibody activity were similar when the same sera were tested by a competition RIA in which group- and subgroup-specific antibodies were adsorbed from the test sera before incubation with 125I-Ad5 hexon of fiber.
Evidence for a Common Mucosal Immunologic System
Authors: McDermott Mark R. , Bienenstock John
Published: December 2022

The origins of immunoglobulin-containing cells in intestinal, respiratory, mammary, and genital tissues were studied in CBA/J female mice by using an adoptive lymphocyte transfer method. Within 24 hr after transfer, [3H]thymidine-labeled donor mesenteric lymph node (MLN) cells were observed in recipient gut, cervix and vagina, uterus, mammary glands, and MLN, where approximately 60% contained IgA and 25% IgG. In peripheral lymph nodes (PLN), 44% of the labeled cells after MLN transfer contained IgG, whereas only 8% were of the IgA isotype. The preference of the MLN to populate mucosal sites was clear from the results. Labeled PLN cells were transferred and the majority of these returned to their sites of origin and contained IgG. Of the small number of labeled PLN cells found in mucosal tissues, approximately equal percentages (30%) of IgA- and IgG-containing cells were seen. Dividing cells prepared from mediastinal (bronchial) lymph nodes (BLN) showed a propensity to localize in the lungs rather than the intestine. However, the predominant immunoglobulin content of these donor cells in gut, lungs, and MLN was IgA. In recipient PLN, most labeled BLN cells contained IgG. These data support the concept of a common mucosal immunologic system.
The Molecular Identification of Two Serologically Defined Gene Products of the Rat Major Histocompatibility Complex
Authors: Natori Takashi , Ohhashi Tatsuhiko , Kotani Tomio , Katagiri Makoto , Aizawa Miki
Published: December 2022

The two SD gene products of the rat major histocompatibility complex were identified by immunochemical means with rat alloantisera. WKA anti-TO antiserum contained anti-TO reactivity possibly detecting a unique private specificity for the Tokyo strain or rats, as well as some other cross-reactivity against common public specificities among ACI, Spd, LEJ, and TO. WKA anti-TO antiserum could react with 125I-labeled AgB4 preparation. The immunoprecipitates gave two peaks of radio-activity on SDS-PAGE corresponding to positions of 37,000 and 12,000 daltons, respectively, showing the basic structure of AgB antigens as described previously. These components were confirmed, by a sequential immunoprecipitation test with rabbit anti-H-2K,D and rabbit anti-HLA-A,B antiserum, to be of the rat major histocompatibility antigens. Two different SD gene products were identified by sequential immunoprecipitation tests in which 125I-labeled AgB4 preparation was first treated with WKA anti-TO antiserum and the residual supernatants were allowed to react with a test antiserum, TO anti-ACI antiserum. Pretreatment of the antigen with WKA anti-TO antiserum could not remove the antigen molecules reactive with TO anti-ACI antiserum. Conversely, pretreatment of the antigen preparation with TO anti-ACI antiserum removed concomitantly the molecules detectable by WKA anti-TO antiserum, suggesting that both molecules carry the same private and/or public specificity(ies). The two SD antigens were inherited together in backcross animals of (ACI × WKA)F1 × WKA, indicating a close linkage of the two SD loci. There was one discrepant animal out of 63 rats tested, suggesting a dissociation of the two loci.
The Separate Roles of Hapten and of Carrier in the Conformation of T Cell-Reactive Determinants on Aba-Protein Conjugates
Authors: Ray Alex , Ben-Sasson Schlomo
Published: December 2022

When new antigenic determinants were formed on a protein by modifying it with p-azobenzenearsonate (ABA), those of the new determinants that were recognized by and activated a T cell-immune response contained functionally separate contributions from the hapten and from the carrier protein. Those parts of the determinant that actually reacted with the T cell recognition site were contributed by the carrier molecule, whereas the hapten contribution was toward specifying the structural arrangement of those parts of the carrier molecule. The indirect nature of the haptenic contribution to the conjugate determinant explains why p-azobenzenearsonic acid on the autologous carrier guinea pig albumin (GPA) cross-reacted with p-azobenzoic acid on the same protein, whereas p-azobenzenearsonic acid in its free form on tyrosine did not cross-react with conjugates of p-azobenzoic acid. Although identity of the hapten components of the autologous albumin conjugates were not necessary for inducing cross-reactive immunity, neither was identity of both hapten and protein moiety sufficient to induce cross-reaction, since ABA-GPA did not induce immunity to ABA-Actyr GPA, and ABA-Actyr-GPA induced only a very weak immunity to ABA-GPA. Thus, the manner or site of attachment of ABA to protein was a major factor in determining the conformation of the determinant that it induced on the protein.

In spite of the finding of some cross-reaction between different protein conjugates of ABA, there was no evidence that the conjugate determinants had induced T cell immunity to any structure containing the hapten in an immunologically recognizable form. ABA-protein immune cells did not respond to ABA-amino acids nor to ABA on the amino acid copolymers GT or GL. Enrichment of the cross-reactive population of ABA-OA or ABA-GPA immune T cells by in vitro selection on macrophage pulsed with the cross-reacting antigen did not amplify any small population of hapten-reactive cells that might have been undetectable by direct testing of an unenriched cell population with ABA-amino acids.
Ultraviolet (UV) Sensitive Leukocyte Allostimulation Cross-Reacts with UV Resistant Sperm
Authors: Levis William R. , Dattner Alan M.
Published: December 2022

Our group has previously reported that human sperm cells are capable of stimulating allogeneic lymphocytes. We now report that the HLA-D on sperm cells is resistant to ultraviolet (UV) light in contrast to HLA-D on leukocytes, which is very sensitive to UV light in the dosage range employed in these experiments. Responder leukocytes were cultured with varying concentrations of stimulating blood leukocytes and varying concentrations of sperm cells washed free of the highly suppressive seminal plasma factor(s). Stimulator blood and sperm cells were exposed simultaneously to UV light delivered from a G.E. cold quartz No. G 15-T8 (2537 Å germicidal) lamp at doses varying from 800 to 25,000 ergs/mm2 as measured by an IL 770 photometer with a narrow band NB254 filter. Even the lowest dosage of UV completely or nearly totally abrogated the stimulating capability of leukocytes whereas sperm cells showed excellent stimulating ability with as much as 25,000 ergs/mm2. We have also been able to clonally prime lymphocytes with sperms. After 10 to 14 days in first or primary culture, sperm-stimulated blast-derived lymphocytes were recultured. The allogeneic responding lymphocytes showed accelerated 3HTdR incorporation to leukocytes from the original sperm donor. We conclude that the UV-resistant HLA-D on sperms share some determinants with the UV-sensitive HLA-D on leukocytes.
Evidence for Shared Idiotypy Expressed by the IgM, IgG, and IgA Serum Proteins of a Patient with a Complex Multiple Paraprotein Disorder
Authors: Hopper John E. , Haren J. Marie , Kmiecik Thomas E.
Published: December 2022

The results of a comparative idiotypic analysis of multiple Ig paraproteins isolated from the serum of an individual patient, Ca, with Sjögren's syndrome and Waldenström's macroglobulinemia are reported. At initial presentation, Ca serum was found to contain two major paraproteins, an IgMκ and an IgGκ, together with a small elevation in the level of IgA protein. The patient's clinical course was characterized by dramatic and opposing changes in the respective serum levels of the IgMκ and IgGκ paraproteins over an extended time period that coincided in part with received chemotherapy. Idiotypic antigenic analysis of the IgMκ and IgGκ paraproteins revealed that the two monotypic proteins shared identical idiotypic determinants. The Ca IgA serum fraction, specifically isolated by an immunoabsorbent and free of any IgG and IgM, was shown to possess idiotypic determinants identical to the IgG and IgM proteins. In extensive tests of specificity, the idiotypic determinants shared by Ca IgM, IgG, and IgA proteins were not present in large excesses of heterologous IgM and IgG, nor on Ig molecules contained in a large number of normal and myeloma sera.
Surface Expression and Synthesis of I-A and I-E/C Encoded Molecules by B Lymphocytes and Ig-Secreting Cells
Authors: Vitetta Ellen S. , Cook Richard G.
Published: December 2022

Splenocytes from recombinant mice were radiolabeled before or after deletion of subpopulations by cytotoxic anterisera (+C) directed against I-A, I-E/C, IgM, or Ig. Examination of the lysates of the surviving cells by immunoprecipitation demonstrated that 1) virtually all I-A and I-E/C molecules are co-expressed and synthesized by Ig+, IgM+ lymphocytes, 2) I-A, I-E/C, and IgM molecules are present on many of the cells secreting IgM and IgG, and 3) populations of Ig-bearing or Ig-secreting cells that lack detectable I-A and I-E/C antigens can be identified in spleen cell populations. The co-expression of I-A and I-E/C on most cells of the B cell lineage is discussed in terms of our present concepts of Ir gene control of immune responses.
In Vitro Analysis of Allogeneic Lymphocyte Interaction
Authors: Delovitch Terry L. , Sohn Uik
Published: December 2022

A helper allogeneic effect factor (AEF) was produced across an incompatibility at the minor histocompatibility loci. This AEF is genetically restricted in its activity since it helps B cells only of the stimulator haplotype and of haplotypes that share both an Mls and I-region identity with the stimulator haplotype. The I-region genes involved here map to the I-A and/or I-C subregions. An anti-LyM immunoadsorbent column but neither an anti-H-2 nor an anti-Ia column adsorbed AEF helper activity. It is suggested that the activation of T helper cells by a positive allogeneic effect across a minor H locus difference and their genetically restricted interaction with allogeneic B cells may in part be due to the acquisition by alloactivated T cells of stimulator cell-derived LyM and/or Mls determinants. The data presented indicate that helper T cells recognize Mls locus alloantigens in the milieu of self Ia antigens.
Functional Analysis of H-2-Linked Genetic Control of Immune Responsiveness to TNP-MSA
Authors: Urba Walter J. , Hildemann William H.
Published: December 2022

The antibody response to trinitrophenyl-conjugated mouse serum albumin (TNP-MSA) in inbred strains of mice is regulated by a gene(s) in the I-B subregion of the H-2 complex. High responsiveness in low H-2k × high H-2b F1 hybrids is inherited as a recessive trait. Various parameters of the immune response to TNP-MSA were investigated to determine whether specific Ir gene activation by TNP-MSA leads to a similar or different type of regulation compared to dominant H-2-linked Ir genes. Analysis of high and low responder sera by radioimmunoassay and isoelectric focusing revealed that Ir-6 controls the production of IgG antibodies in high responder mice. Although both high and low responder immune sera contained equivalent amounts of IgM TNP-antibody, IgG anti-TNP activity was found only in high responder sera. However, low responders could be induced to produce high levels of IgG TNP-antibody after immunization with TNP-MSA complexed to a xenogeneic carrier (MBSA). Adult thymectomy and rabbit antimouse T cell serum treatment of high and low responder animals demonstrated that TNP-MSA is capable of eliciting both a T-independent IgM response and a T-dependent IgG response. The thymus-dependent IgG produced by high responder animals is of restricted heterogeneity as revealed by isoelectric focusing. High responsiveness to TNP-MSA displays either dominant or recessive inheritance in F1 progeny depending on the H-2 haplotype combination. This finding is interpreted in terms of Ir-6 gene product interaction and hierarchical or partial dominance among multiple alleles.
Murine Antibody-Dependent Cellular Cytotoxicity to Herpes Simplex Virus-Infected Target Cells
Authors: Kohl Steve , Cahall David L. , Walters Deborah L. , Schaffner Vann E.
Published: December 2022

Freshly collected peritoneal cells (PC) and cultured spleen cells (SC) (but not fresh SC) from nonimmune mice could mediate antibody-dependent cellular cytotoxicity (ADCC) against herpes simplex virus (HSV)-infected cells in the presence of mouse or human sera containing antibody to HSV. PC also demonstrated variable natural killer cell cytotoxicity to infected cells.

Both PC and cultured SC required high concentrations of antibody and high effector to target cell ratios for optimal ADCC. The time kinetics of the reaction appeared to depend on the state of activation of the effector cells. In both PC and SC populations, ADCC activity was limited to adherent cells, and was profoundly inhibited by particulate latex or silica.

The murine effector cell found in PC and SC able to mediate ADCC to HSV-infected cells appears to be a macrophage.
Gene Dose Effects in Ir Gene-Controlled Systems
Authors: Dorf Martin E. , Stimpfling Jack H. , Benacerraf Baruj
Published: December 2022

(B10.A × B10.S)F1 hybrid mice produce lower levels of anti-GLφ antibody than B10.S(9R) and B10.HTT mice. To determine whether this difference is due to a gene dose or a cis-trans effect, [B10.S(9R) × B10.S(8R)]F1 mice were immunized with GLφ. These mice carry one dose of the responder Ir gene alleles in the cis position whereas the recombinant B10.S(9R) and B10.HTT mice carry two doses of the relevant genes. Both (B10.A × B10.S) and [B10.S(9R) × B10.S(8R)] F1 mice produced comparably lower amounts of antibodies as compared to the recombinant strains. The data therefore demonstrate that gene dose and not cis-trans effect accounts for the differences between F1 and recombinant strains in their antibody response to GLφ controlled by complementary Ir genes.
The LT System in Experimental Animals
Authors: Hiserodt John C. , Tiangco Gerry J. , Granger Gale A.
Published: December 2022

Lymphocytes or purified T cells obtained from the spleens of alloimmune C57BL/6, DBA/2, or C3H/DiSn mice, when placed on monolayers of lectin-coated allogeneic (L-929 or 3T3) fibroblasts, release into the supernatant various forms of cell lytic material. One form appears to be a high m.w. complex containing an antigen-binding receptor(s) that is highly labile and capable of causing rapid and specific lysis of allogeneic target cells in vitro. Material(s) that could mediate these cell lytic effects were detected in culture supernatants as early as 3 hr after stimulation, peaked at 6 to 9 hr, and declined thereafter. The specific cell lytic activity appeared to be due to high m.w. LT-receptor complexes for the following reasons: a) antisera that could neutralize murine LT activity in vitro could inhibit this effect; b) absorption of supernatants on the specific target cells at 4°C removed both the specific lytic activity and nonspecific LT activity detectable on L-929 cells in vitro; c) this material(s) was highly unstable, as are LT complex forms; and d) fractionation by molecular sieving of these supernatants revealed that cell lysis was mediated by material(s) in the high (>200,000) m.w. complex form(s). The lytic effect did not appear to be due to Ab + C because supernatants that had lost their specific lytic activity could not be reconstituted with fresh sources of C. Since purified alloimmune T lymphocytes yielded more active supernatants than unseparated nonadherent spleen cells, and polyspecific goat anti-mouse Ig sera had virtually no effect on this lytic activity, we feel the receptor(s) in these complexes originates from the T cell(s). The data support the concept that the short-lived specific cell lytic material in these supernatants is a high m.w. complex containing αH m.w. LT subunits in functional association with specific (T cell?) antigen-binding receptor(s) molecules.

These findings strongly corroborate analogous findings in the human and support the concept that the smaller m.w. LT molecules represent a system of weakly lytic but related subunits released by cells that can associate together and functionally associate with antigen-binding receptor(s) to form highly effective cell lytic complexes. Furthermore, these lytic LT-receptor complexes can be directed by the specificity of the receptor with which they are associated.
Use of a Monoclonal Antibody (W6/32) in Structural Studies of HLA-A,B,C Antigens
Authors: Parham Peter , Barnstable Colin J. , Bodmer Walter F.
Published: December 2022

The experiments reported here show that W6/32 antigenic activity is retained in papain-solubilized HLA antigens and that the W6/32 antibody provides a useful standard reagent to detect and assay HLA-A,B,C antigens. The W6/32 antibody was purified and used to construct an immunoaffinity column. Soluble HLA-A,B,C antigens from papain or detergent-treated membranes could be highly purified in a single step with this column and serologic analysis showed that HLA-A,B and C antigens were bound to the column. Thus, the W6/32 antigenic determinant is present on gene products of all three loci.

The amount of HLA-A,B,C antigens on the surface of human B cell lines and peripheral blood lymphocytes was measured with W6/32 antibody. B cell lines expressed, on average, 9 times as much cell surface HLA-A,B,C antigens as peripheral lymphocytes, although there was appreciable variation within each group. The B cell line Bri 8, for example, expressed 1.5 × 106 W6/32 antigenic sites, and by inference, HLA-A,B,C molecules per cell. Equal amounts of β2m and HLA-A,B,C chain were found on both cell types.

The isolated HLA-A chain from intact 125I-HLA-A2 antigens weakly bound to W6/32 antibody in contrast to 125I-β2-microglobulin (β2m) isolated from the same preparation of HLA-A2 antigens that showed no demonstrable binding. when an excess of cold β2m was added to the isolated 125I-HLA-A2 chain, the binding to W6/32 antibody was considerably enhanced. These results suggest that the W6/32 antigenic determinant involves only amino acids of the HLA-A,B,C chain and is a product of their three dimensional configuration. Stable maintenance of this configuration appears to be dependent on the association of the HLA-A,B,C chain with β2m.
Studies on in vivo Priming of the TNP-Reactive Cytotoxic Effector Cell System
Authors: Fujiwara Hiromi , Levy Robert B. , Shearer Gene M. , Terry William D.
Published: December 2022

A comparison was made of the effects of i.v. inoculation of trinitrophenyl-(TNP) conjugated syngeneic cells on the subsequent in vitro generation of TNP-reactive effector cell activity and on the in vivo development of TNP-contact sensitivity. The administration of syngeneic TNP-conjugated spleen cells before 2,4,6-trinitro-1-chlorobenzene (TNCB) painting abolished the capability of animals to develop TNP-contact sensitivity. In contrast, the same treatment resulted in an appreciable augmentation in the generation of TNP-reactive cytotoxic effector cell activity as measured by subsequent in vitro sensitization with TNP-conjugated cells. The possible mechanisms by which enhanced TNP-reactive cytotoxic effector cell activity was elicited under conditions identical to those that induced unresponsiveness for TNP-contact sensitivity are discussed.
Activation of Human Natural Killer Cells Cytotoxic for Human Leukemia Cells by Purified Interferon
Authors: Zarling Joyce M. , Eskra Linda , Borden Ernest C. , Horoszewicz Julius , Carter William A.
Published: December 2022

Interferon (IF) inducers and IF-containing preparations of leukocyte origin have recently been reported to augment natural killer (NK) cell activity. Our studies were undertaken to determine whether highly purified human fibroblast interferon (HFIF), induced by polyinosinic: polycytidylic acid (poly I:C), would augment human NK cell activity against an NK-sensitive leukemia cell line, K562, and, more importantly, to determine whether HFIF would activate normal lymphocytes to lyse human leukemia cells that have not been adapted to tissue culture or established as cell lines. Pretreatment of peripheral blood mononuclear cells for 1 to 16 hr with purified HFIF (150 to 300 units/ml) caused 2.5- to 5-fold increases in cytotoxicity against the K562 cells. Both native and HFIF-augmented cytotoxicities were mediated by FcR+ lymphocytes (lymphocytes expressing receptors for the Fc portion of IgG). Removal of FcR+ lymphocytes, but not monocytes, prevented HFIF from augmenting cytotoxicity, suggesting that HFIF activates preexisting FcR+ NK cells and that monocytes are not required for this effect. Our findings that NK cell activity is augmented by poly I:C-induced fibroblast IF, which lacks lymphokines and viral products present in virus-induced leukocyte IF preparations previously used, together with the fact that our IF preparation was more highly purified than any previously used for such studies, strongly suggest that the IF protein itself augments NK activity.

HFIF-treated cells from some individuals lysed many patients' leukemia cells; however, the HFIF-treated cells were not cytotoxic for autologous or allogeneic normal lymphocytes or concanavalin A blasts. FcR+ cells were required for the HFIF-induced cytotoxicity against the patients' leukemia cells. These results thus demonstrate that not only leukemia cell lines, but also human leukemia cells that have not been adapted to tissue culture or established as cell lines, can be lysed by IF-activated human NK cells.
Studies on the Terminal Stages of Immune Hemolysis
Authors: Boyle Michael D. P. , Borsos Tibor
Published: December 2022

The inhibitory effects of 0.1 M EDTA on the lysis of E* prepared by incubating EA with whole GPC was studied. At high end point lysis (>70%) 0.1 M EDTA failed to prevent hemoglobin release whereas at lower end point (<60%) 0.1 M EDTA was effective. In all cases hemoglobin release was inhibited by 25% BSA. When E* were prepared by incubating EAC1-8 with C9, similar results were obtained. In this system the difference in the ability of 0.1 M EDTA to inhibit hemoglobin release at high or low end point lysis could not be correlated with the number of lesions/cell but appeared to be related to the C9 to SAC1-8 ratio. With limiting SAC1-8 and excess C9, E* were produced from which hemoglobin release could not be prevented by 0.1 M EDTA whereas at lower C9 to SAC1-8 ratios hemoglobin release was prevented by 0.1 M EDTA. These differences most probably reflect functionally different sized transmembrane channels that were produced at different C9 to SAC1-8 ratios.
Studies on the Terminal Stages of Immune Hemolysis
Authors: Boyle Michael D. P. , Gee Adrian P. , Borsos Tibor
Published: December 2022

We have previously shown that 0.1 M EDTA could be used to distinguish functionally different transmembrane channels produced during complement-(C) mediated hemolysis of E. In this paper we have studied the ability of sugars of varying Stokes' radii to prevent hemoglobin release from E* intermediates whose lysis was inhibitable or not inhibitable by EDTA. On the basis of these experiments we propose that the inhibition of E* transformation by high molarity EDTA occurs by virtue of the size of the EDTA molecule in solution. Studies on the effect of EDTA on red cell lysis induced by polyene antibiotics that form transmembrane channels of a defined size support this conclusion. The results of these experiments were interpreted to mean: 1) The EDTA inhibitable lesion on E* has a smaller effective radius than the noninhibitable lesion; 2) the effective radius of the smallest lesion that yields a lytic site was less than 3.6 Å; 3) the lesions produced in the red cell membrane by C are not uniform but vary in size depending on the C9 to SACI-8 ratio used to produce E*.
Latent Lymphokines: Isolation of Guinea Pig Latent Lymphocyte-Derived Chemotactic Factor for Monocytes. Its Activation by Trypsin and a Soluble Factor from Macrophages
Authors: Postlethwaite Arnold E. , Kang Andrew H.
Published: December 2022

Guinea pig lymphocytes when depleted of macrophages and stimulated by the T cell mitogen phytohemagglutinin produce a latent form of lymphocyte-derived chemotactic factor for monocytes (LDCF-M). Latent LDCF-M is also produced when immune lymphocytes are stimulated in vitro with specific antigen, horseradish peroxidase. Latent LDCF-M from both sources can be activated and converted to “classical” LDCF-M by trypsin and by a soluble factor obtained from sonicated macrophages. These observations suggest that macrophages may modulate lymphokine activities in vivo by releasing soluble factors that convert inactive latent lymphokines to biologically active substances.
Specificity and Suppressor Function of Human T Cells Responsive to Autologous Non-T Cells
Authors: Sakane Tsuyoshi , Green Ira
Published: December 2022

Human T cells proliferate in response to autologous non-T cells. This reaction, called the autologous MLR has been shown to have both immunologic specificity and memory. Our present study first demonstrates that specific secondary responses can be demonstrated in the autologous MLR. A BUdR and light technique was then used to ablate cells proliferating in response to either autologous or allogeneic non-T cells. The removal of T cells responsive to autologous non-T cells left the responsiveness to allogeneic T cells relatively intact, and conversely the removal of T cells responsive to allogeneic non-T cells left the responsiveness to autologous non-T cells relatively intact. Thus, the T cells responsive to autologous non-T cells and those responsive to allogeneic non-T cells are largely separate cell populations.

Since the lymphocytes from patients with SLE fail to demonstrate a normal autologous MLR and also have a defect in the development of Con A-induced suppressor cells, we investigated whether the T cell population responsive in the autologous MLR was particularly enriched in cells capable of developing suppressor capacity. A BUdR and light treatment was used to remove T cells responsive to either autologous or allogeneic non-T cells. The remaining cells were then activated by Con A, and their subsequent suppressor activity was measured. Removal of T cells proliferating in response to autologous non-T cells produced significantly greater failure of suppressor cell development than did the removal of T cells responsive to allogeneic non-T cells. Thus, T cells capable of responding to autologous non-T cells are particularly enriched in cells capable of becoming suppressor cells.
Human Monocyte, Lymphocyte, and Granulocyte Antibody-Dependent Cell-Mediated Cytotoxicity Toward Tumor Cells
Authors: Levy Paul C. , Shaw George M. , LoBuglio Albert F.
Published: December 2022

Previous investigations of antibody-dependent cell-mediated cytotoxicity (ADCC) by human leukocytes indicated that lymphocytes and granulocytes, but not monocytes, lysed tumor cell targets. We have recently shown that monocytes, too, can carry out ADCC toward at least three allogeneic tumor cell lines. This paper examines the characteristics of ADCC by all three Fc receptor-bearing human leukocytes (monocytes, lymphocytes, and granulocytes) toward the same antibody-coated 51Cr-labeled tumor target cells. For each leukocyte effector, ADCC was found to be temperature dependent and rapid, with most target cell lysis occurring within 4 hr of incubation. The extent of target cell destruction was positively related to the effector cell/target cell ratio and to the degree of antibody sensitization of the target cells. ADCC was clearly antibody dependent since the cytotoxic event did not involve nonantibody-coated “innocent bystander” target cells and was inhibited by staphylococcal Protein A and nonspecific monomeric immunoglobulin. Granulocyte ADCC required considerably larger amounts of target cell-bound antibody than did ADCC by lymphocytes and monocytes. Conversely, larger amounts of monomeric immunoglobulin were required to inhibit granulocyte ADCC as compared to monocyte and lymphocyte ADCC in experiments utilizing identical antibody-coated target cells. In innocent by-stander experiments, no evidence for a soluble mediator of target cell lysis was found for any of the effector leukocytes. We suggest that this experimental system which involves three Fc receptor-bearing human leukocyte populations and identical antibody-coated target cell preparations provides a new and unique method for studying this cell-mediated cytotoxic event.
The Role of H-2-Linked Genes in Helper T Cell Function
Authors: Swierkosz James E. , Marrack Philippa , Kappler John W.
Published: December 2022

We have studied the H-2 genes that control interactions between antigen-presenting macrophages (Mø) and helper T cells both during and after priming. B6AF1 (H-2b × H-2a) mice were initially primed in vivo with keyhole limpet hemocyanin (KLH)-pulsed Mø of either parental H-2 type, and their T cells were tested for helper activity in the in vitro anti-trinitrophenol (TNP) response to TNP-KLH. In each case, F1 T cells cooperated well only with B cells and Mø having the same K-I-A subregion as the Mø used for priming.

In a second approach, F1 mice were instead primed with free antigen, and T cells enriched in specific helper function were then isolated in vitro by adherence to antigen-pulsed parental Mø. The F1 T cells isolated in this manner exhibited enriched helper activity only when tested with B cells and Mø that had the K-I-A region type of the Mø used for isolation. With this technique, the same genetic restrictions were also found for responses to sheep red blood cells (SRBC).

A direct role for I-region genes in this system was established by the ability of anti-Ia alloantisera to specifically inhibit the enrichment of helper T cells on Mø monolayers. Taken together these results pinpoint I-A as the relevant subregion involved in the associative recognition of these complex antigens by helper T cells.
Autoimmune Cell-Mediated Responses: Spontaneous Responses to Syngeneic Cells in NZB/NZW F1 Mice
Authors: Senitzer David , Cafruny William , Raeder Roberta , Freimer Earl H.
Published: December 2022

The effect of various stimulator cell populations on B/W mouse spleen cell cultures was analyzed by measuring 3H-thymidine incorporation. The data show that syngeneic heart and kidney cells induce strong stimulation of DNA synthesis in B/W spleen cells. In contrast, syngeneic liver cells are not stimulatory. Autologous stimulation of adult B/W spleen cells by heart and kidney cells was also observed. BALB/c heart and kidney cells were stimulatory for B/W spleen cells, although BALB/c spleen cells were only slightly stimulated by B/W or BALB/c heart and kidney cells.

The responses to syngeneic heart and kidney cells were observed to peak at 7 to 8 days of culture. The magnitude of the observed responses did not appear to correlate with the age or sex of the B/W spleen cells used, and the ability to response to heart and kidney cells with large increases in DNA synthesis occurred at a very early age. Spleen cells from old B/W mice lose the ability to become stimulated by concanavalin A, although no concomitant loss of reactivity to syngeneic heart or kidney cells is observed. The presence of θ-bearing lymphocytes in B/W spleen cell populations is not required during culture initiation for the stimulation of DNA synthesis by syngeneic heart or kidney cells. However, during the peak of DNA synthesis, the majority of responder spleen cells carry θ-antigen.

We postulate that antigenic determinants present on heart and kidney cells induce in vitro cell-mediated immune responses of an autoimmune nature in B/W spleen cell populations. The results are in aggrement with an early defect in immunologic regulation that persists throughout the life of the B/W mouse.
Cloning of B Cells Positive or Negative for Surface IgD
Authors: Layton Judith E. , Teale Judy M. , Nossal G. J. V.
Published: December 2022

As in the accompanying paper, spleen cells from 2½-week-old mice, containing equal proportions of sIgD+ and sIgD- B cells were separated in the fluorescence-activated cell sorter into sIgD+ and sIgD- populations. The activity of each population and the effect of tolerance induction were measured in a T-dependent B cell-cloning assay: the splenic focus assay of Klinman and colleagues. The anti-DNP precursor frequency was found to be essentially the same in each population when calculated on a per B cell basis. In addition, the response from the sIgD- population was unlikely to be from cells that had acquired sIgD during the culture period, since the presence of anti-δ serum in the cultures did not inhibit the response from this population. The isotype of the antibody produced by responding clones was analyzed and the results showed that the sIgD- population produced a slightly higher proportion of clones secreting IgM only, and a lower proportion of IgG-secreting clones, than the sIgD+ population. Tolerance induction in the separated populations indicated that the sIgD- subset was only marginally susceptible to tolerogenesis whereas the sIgD+ subset was resistant. Furthermore, if the expected contribution of the highly tolerizable sIg- cells contained in the sIgD- subset was subtracted, the degree of tolerance was reduced to an insignificant level. Thus, the response to a T-dependent antigen is not restricted to the sIgD+ subset of cells and resistance to tolerogenesis does not appear to be caused by acquisition of sIgD, even when a T-dependent challenge antigen is used. The results are discussed in the light of other work on the function of IgD.
Nature of T Lymphocyte Recognition of Macrophage-Associated Antigens
Authors: Thomas David W. , Meltz Susan K. , Wilner George D.
Published: December 2022

The immune response of strain 2, strain 13, and (2 × 13)F1 guinea pigs was studied after immunization with human fibrinopeptide Bβ(hFPB), a 14 amino acid (Bβ1-14) thrombin-derived fragment of fibrinogen. Several weeks after immunization, strain 2 and (2 × 13)F1 antimals showed a positive anti-hFPB delayed-type hypersensitivity skin test and produced a dramatic hFPB-specific T lymphocyte proliferative response in vitro, as assessed by the incorporation of 3H-thymidine. Strain 13 animals immunized with hFPB did not produce an anti-hFPB skin test and showed little or no in vitro T cell proliferation with hFPB. Neither guinea pig strain produced detectable anti-hFPB antibody as determined by radioimmunoassay. Immune T cells were also tested for their ability to respond to several sequential synthetic homologues of hFPB. Strain 2 and (2 × 13)F1 T cells immunized with hFPB (Bβ1-14) were stimulated by fragments Bβ3–14, 5–14 and 7–14, but not by fragments Bβ1–13 and 9–14. In addition, strain 2 and strain 13 guinea pigs were immunized with the various homologues themselves and the immune T cells tested for responsiveness in vitro with hFPB and a battery of the fragments. Strain 2 T cells immunized with Bβ1–13 showed no proliferative response with Bβ1–14, 3–14, 5–14 and 7–14, and neglible stimulation with the homologous antigen Bβ1–13. After immunization with Bβ5–14 and 7–14, strain 2 T cells showed a proliferative response with Bβ1–14, 3–14, 5–14, and 7–14, but not with Bβ1–13. T cells from strain 2 animals immunized with B 9–14 were unresponsive with all homologues tested. T cells from strain 13 guinea pigs immunized with Bβ1–13 produced a dramatic proliferative response with Bβ1–13, but were unresponsive with Bβ1–14, 3–14, 5–14, and 7–14. Strain 13 animals immunized with Bβ5–14 and 9–14 showed no T cell response with any of the homologues tested, whereas Bβ7–14 immune strain 13 T cells produced a small response with Bβ7–14. These results are discussed with respect to T cell recognition of specific amino acid sequences of the fibrinopeptide molecule.
Ia Antigens in Mouse Skin Are Predominantly Expressed on Langerhans Cells
Authors: Tamaki Kunihiko , Stingl George , Gullino Marisa , Sachs David H. , Katz Stephen I.
Published: December 2022

We have investigated the expression of products of the mouse major histocompatibility complex (MHC) on BALB/c and A/J epidermal cells. By using reagents with specificity for various products of the MHC in an indirect immunofluorescence procedure, we found that H-2 antigens are expressed on the vast majority of epidermal cells. Ia antigens, by contrast, are present on only 2.4 to 6.9% of all epidermal cells. These Ia-bearing cells bear a receptor for the Fc portion of IgG and ultrastructurally exhibit the characteristics of Langerhans cells. Ia antigens on Langerhans cells are encoded for by at least the I-A and I-E/C subregions of the MHC.
Active Disassembly of the First Complement Component, C1̄, by C1̄ Inactivator
Authors: Ziccardi Robert J. , Cooper Neil R.
Published: December 2022

In order to clarify its mechanism of disassembly, the first component of human complement, C1, was reconstituted from C1q, 125I-C1r, and 131I-C1s. This radiolabeled C1 bound tightly to aggregated IgG covalently linked to Sepharose 4B. The subsequent addition of C1̄ inactivator (C1̄-In) led to the rapid release of equimolar quantitites of 125I-C1r̅ and 131I-C1s̅ from the agg IgG. SDS-PAGE analyses of such eluates showed that one C1̄-In molecule was associated with each C1r̅ polypeptide chain and another C1̄-In molecule was bound to each C1s̅ polypeptide chain. The released C1r̅, C1s̅, and C1̄-In co-sedimented in sucrose density gradients with a rate of 9S and monospecific anti-C1s̅ pelleted all three proteins. Therefore, C1r̅, C1s̅, and C1̄-In, in a molar ratio of 1:1:2, respectively, are released from C1̄ as a complex.

An analogous 9S complex containing C1r̅, C1s̅, and C1̄-In was generated in normal human serum after incubation with activators of the classical complement pathway. The C1r̅C1s̅(C1̄-In) complexes generated in both serum and the purified system were stable in the presence of EDTA.

A diffusion coefficient of 2.3 × 10-7 cm2/sec was determined for the C1r̅C1s̅(C1̄-In) complex from its behavior on gel filtration. An m.w. of 330,000 was then calculated from its sedimentation and diffusion coefficients. This m.w. is consistent with the value of 382,000 for a molecule of composition C1r̅C1s̅(C1̄-In)2 obtained by summing the weights of the subunits. These results indicate that C1-In efficiently disassembles C1̄, thereby releasing two C1r̅C1s̅(C1̄-In)2 complexes per C1̄ molecule.
Sequentially Derived Mutants of the Constant Region of the Heavy Chain of Murine Immunoglobulins
Authors: Morrison Sherie L.
Published: December 2022

In an effort to define the mechanism of the γ2b → γ2a subclass switch observed in cultured cells, a series of constant region mutants has been isolated in the cell line (45.6) derived from the MPC-11 tumor. The investigation was begun with 10-1, a spontaneous variant of 45.6 which synthesizes a 47,000 dalton heavy chain with a deletion in CH1. From 10-1, two types of secondary variant was synthesizing a 55,000 dalton heavy chain that was γ2a serologically. Five independent variants synthesizing 55,000 dalton γ2a heavy chain were isolated from 10-1. All five were of the same apparent m.w. and four, but not the fifth, had identical isoelectric focusing patterns. The second type of secondary variant isolated from 10-1 was synthesizing a heavy chain even smaller than the heavy chain of 10-1. Two different variants of this type, I17 and G251, synthesized heavy chains of approximately 35,000 and 40,000 daltons, respectively, which did not react with any subclass-specific antiserum.

From I17 and G251 it was possible to isolate tertiary variants that synthesized γ2a heavy chains of either 47,000 daltons or 55,000 daltons m.w. The 55,000 dalton γ2a heavy chains exhibited many different isoelectric focusing patterns. Peptide map analysis of two 47,000 dalton and two 55,000 dalton γ2a tertiary variants showed no differences from the Fc of MOPC-173, a naturally occurring γ2a myeloma. However, a third 55,000 dalton γ2a variant synthesized a heavy chain with a Fc that was apparently not identical to that of MOPC-173.

These experiments show that it is possible to isolate γ2b → γ2a subclass switch mutants from variants synthesizing short heavy chains. It is not possible to define unequivocally the mechanism of the subclass switch. However, it clearly does not result from the translocation of an intact V region to an intact constant region. The resulting data are, however, compatible with the γ2b → γ2a switch resulting from a recombination event between tandem genes on homologous chromosomes or chromatids.