bluuebunny/crossref_metadata_2025 huggingface dataset:
Biologic Activities of Rabbit Immunoglobulin G in Relation to Domains of the Fc Region
Authors: Ovary Zoltan , Saluk Paul H. , Quijada Luis , Lamm Michael E.
Published: December 2022

Biologic properties of antibodies are known to be mediated by the Fc portion of the H chains. In the present study such properties as complement fixation, cutaneous anaphylaxis, and macrophage cytophilia were examined in relation to the CH2 and CH3 domains of rabbit IgG. Fragments containing but one of these domains were prepared from plasmin and papain digests. Facb fragments of anti-DNP antibodies, together with the antigen DNP-BSA, were able to fix complement by the classical pathway, a result which implicates the CH2 domain; however, guinea pig Fab fragments directed to regions of the rabbit antibody molecule other than CH2 were able to inhibit complement fixation. Facb fragments were unable to mediate PCA or reverse PCA reactions in guinea pigs, nor were CH3 fragments active in tests of reverse PCA or inhibition of PCA. These results suggest that the entire Fc region is needed for cutaneous anaphylaxis. The ability to bind to guinea pig lung macrophages was studied with a rosette technique. Facb fragments were active whereas CH3 fragments failed to inhibit. It is suggested that although some effector functions of antibodies can be assigned to individual domains, others require the entire Fc region.
Release of Tumor-Associated Antigens by Murine Melanoma Cells
Authors: Bystryn Jean-Claude
Published: December 2022

Synthesis and release of radiolabeled macromolecules and tumor-associated antigens (MAA) by murine B16 melanoma was studied by pulse labeling cells in culture with 3H-leucine. Approximately 36% of newly synthesized macromolecules and 44% of newly synthesized MAA were released in 48 hr. MAA release was slightly, but consistently, more rapid than the average release of other macromolecules. Release of MAA did not result solely from cell death since it was greater than that of 51Cr-labeled molecules and cell viability was over 98%. The rate of release of newly synthesized MAA was not significantly influenced by cell replication. However, synthesis of MAA was much greater during the logarithmic than the stationary phase of cell growth, suggesting a concomitant increase in the amount of MAA available for release.

These findings indicate that antigens and other macromolecules can be rapidly released by viable tumor cells.
Enumeration and Isolation of Rabbit T and B Lymphocytes by Using Antibody-Coated Erythrocytes
Authors: Wilson Barry S. , Teodorescu Marius , Dray Sheldon
Published: December 2022

Rosette formation with antibody-coated erythrocytes (Ab-E) was employed for the enumeration and isolation of rabbit B cells (Ig+T-) and T cells (Ig-T+). The cells bearing surface Ig (Ig+ cells) were enumerated by a direct immunocytoadhesion technique utilizing anti-rabbit IgG antibody-coated erythrocytes (Ab-E). To enumerate cells bearing thymus cell antigen (T+ cells), an indirect rosette technique was used in which lymphocytes were first sensitized with guinea pig anti-rabbit thymus cell antiserum and then rosetted with anti-guinea pig IgG Ab-E. To demonstrate the specificity of the anti-thymus cell antiserum, a 51Cr radioimmunoassay for counting rosettes was employed along with visual counting to enumerate Ig+ and T+ cells in lymph node cell populations. When Ig+ and T+ lymph node cells were rosetted simultaneously with sheep and human erythrocytes, no mixed rosettes (less than 1%) were observed. Ficoll-Hypaque gradient centrifugation was used to obtain purified Ig+T- and Ig-T+ cells by removing rosetted T+ and Ig+ cells, respectively. The purity of isolated Ig-T+ cells was indicated by 94 to 95% indirect rosetting with anti-thymus cell antiserum and by 0 to 3% direct rosetting with anti-rabbit IgG Ab-E. The purity of isolated Ig+T- cells was indicated by 90 to 94% direct rosetting with anti-rabbit IgG Ab-E and by 2 to 3% indirect rosetting with anti-thymus cell antiserum. The percentages of Ig+T- and Ig-T+ cells were determined in peripheral blood and in various lymphoid organs. The isolated Ig+T- and Ig-T+ cells were also characterized by their responses to mitogens. Thus, nearly pure Ig+T- and Ig-T+ cells were isolated by “negative selection,” which should minimize functional changes of the cells, and thereby facilitate the study of their biologic properties, e.g., their response to mitogens.
Pathogenetic Mechanisms in Immune Polioencephalomyelitis: Quantitative Evaluation of Protective and Pathogenetic Effects of Lymphoid Cells
Authors: Duffey P. S. , Lukasewycz O. A. , Martinez D. , Murphy W. H.
Published: December 2022

Terminal dilution, adoptive cell transfer techniques were developed to quantify the protective effect of lymphoid cells in the pathogenesis of immune polioencephalomyelitis (IPE). The pathogenetic effects of lymphoid cell populations were quantified by deleting the step of antigenic challenge. Regression curves were computer analyzed and PD50 values were compared. Immune spleen cells (ISC) from 4- to 6-week-old donors were more protective (PD50 = 4.9 ± 1.3) than ISC from 12-month-old animals (PD50 > 7.0). The slopes of the regression curves also differed markedly (young mice, - 0.24; old mice, - 0.09). ISC were less protective in 12-month-indicator mice than in 5-month-old recipients (PD50 values of 5.2 ± 0.8 and 3.7 ± 0.8, respectively). When adoptive cell transfer tests were used to quantify the pathogenetic effects of donor cells it was found that ISC were pathogenetic at doses of 105 or less, but protective at higher doses. IPEC were pathogenetic at all test doses. When ISC were x-irradiated or sonicated they were only pathogenetic. Normal spleen or peritoneal exudate cells were neither protective nor pathogenetic.

A model was developed in which mice were either thymectomized mectomized at birth (Tx), or Tx at birth and x-irradiated (500 R) 8 weeks later (Tx-XR). Sham Tx or Tx-XR mice served as controls. All of the mice were challenged with antigen (104 x-irradiated Ib cells). Only a portion (8/24) of the Tx mice developed IPE, indicating that resistance was T cell dependent but also involved a significant T cell independent component. The data also indicated that T cells were not pathogenetic effector cells in this model. Tx mice were not reconstituted by ISC (7/18 developed IPE), Tx-XR mice were partially reconstituted (3/12 developed IPE), but sham Tx-XR were fully restored (0/20 had IPE). Normal spleen cells did not reconstitute any of the mice.
Suppressor Cells in the Spleens of Tumor-Bearing Mice: Enrichment by Centrifugation on Hypaque-Ficoll and Characterization of the Suppressor Population
Authors: Pope Barbara L. , Whitney Richard B. , Levy Julia G. , Kilburn Douglas G.
Published: December 2022

Spleen cells from mice bearing methylcholanthrene-induced sarcomas or a mammary adenocarcinoma suppressed the mitogen responses of normal spleen and lymph node cells. Lymph node cells from tumor bearers had no suppressive effects. Centrifugation of spleen cells layered on Hypaque-Ficoll (specific gravity of 1.08) produced a dense fraction which pelleted and a light fraction which was retained at the Hypaque-Ficoll-medium interface. Suppressive activity was not found in either fraction of normal spleen cells. In tumor-bearer spleen cells suppressor activity was greatly enriched in the light fraction.

Treatment of the suppressor fraction with anti-ϑ or anti-Ig serum and complement did not remove suppressor activity. However, the suppressor cells were removed by passage through nylon wool or by carbonyl iron treatment. Also, the population which adhered to plastic Petri dishes contained the suppressor cell activity.
Immunologic Cross-Reactivity of Antigen(s) Induced by Drug Treatment in Two Leukemic Sublines
Authors: Nicolin Angelo , Bini Alfredo , Di Padova Franco , Goldin Abraham
Published: December 2022

Previous studies have demonstrated that new antigenic specificities, not detectable on parental cells, can be induced by in vivo treatment of murine leukemic cells with anti-neoplastic agents.

The immunologic properties of leukemic cells altered by treatment with 5-(3,3-dimethyl-1-triazeno) imidazole-4-carboxamide (DIC) were investigated further. Immunologic cross-reactivity between two DIC-treated leukemic sublines has been demonstrated by cell-mediated immunity in vitro and by active or adoptive immunity in vivo. Rabbit antiserum to DIC-treated sublines absorbed with the parental cells showed residual activity against the DIC-sublines that was specifically inhibited by further absorption with DIC-cells.
Isolation and Partial Characterization of SAA—An Amyloid-Related Protein from Human Serum
Authors: Rosenthal C. Julian , Franklin Edward C. , Frangione Blas , Greenspan Joel
Published: December 2022

A 12,000 dalton serum amyloid A protein (SAA) has been isolated by chromatography on Sephadex in 10% formic acid. It is similar immunologically to the previously characterized 8500 dalton tissue amyloid A (AA) protein. The results of amino acid analyses, peptide maps, and the identity of the first 11 residues of the SAA and AA proteins support the idea that AA represents the amino terminal fragment of SAA and is derived from it by proteolysis.
Immunoglobulin-Positive Mononuclear Cells in Human Peripheral Blood: Detection by Mixed Anti-Globulin and Direct Anti-Globulin-Rosetting Reactions
Authors: Haegert David G. , Coombs Robin R. A.
Published: December 2022

Ig-bearing mononuclear cells were identified in Ficoll-Hypaque preparations of human peripheral blood by using mixed anti-globulin (MAG) and direct anti-globulin rosettes; indicator cells consisted of sheep erythrocytes coated with human F(ab′)2 or anti-F(ab′)2 antibody, respectively. Of the cell populations isolated from 10 normal subjects, a mean of 68% was lymphocytes. However, fewer than 50% of the cells with detectable surface Ig were lymphocytes. On viable cell preparations using chromic chloride-treated sheep erythrocytes (CrCl3SRBC) coated with anti-F(ab′)2 antibody, a mean of 20.1% of the lymphocytes formed rosettes, i.e., were B. Up to 6% of peripheral blood lymphocytes formed mixed Ig-rosettes and E-rosettes.

On viable lymphocytes using F(ab′)2-coated CrCl3 SRBC, MAG rosettes were insensitive in detection of B lymphocytes. Formaldehyde treatment of lymphocytes increased the number of B cells detectable to 25.5% of the lymphocyte population. Study of T-enriched and B-enriched populations showed that the observed increase in B cell reactivity was real and not due to MAG-rosetting T cells. A one-stage procedure for T and B lymphocyte separation is described.
Redistribution of the Fc Receptor on Human Blood Monocytes and Peritoneal Macrophages Induced by Immunoglobulin G-Sensitized Erythrocytes
Authors: Romans D. G. , Pinteric L. , Falk R. E. , Dorrington K. J.
Published: December 2022

The Fc receptor is a plasma membrane component exhibiting an affinity for the Cγ3 domain of certain subclasses of immunoglobulin G. Using anti-Rho (D)-sensitized red cells (EA) in a slide rosette technique, we have demonstrated a translational mobility and polar redistribution of this receptor on the human blood monocyte and peritoneal macrophage. These cells, isolated from venous blood and malignant ascites and identified histochemically, showed a time- and temperature-dependent receptor capping defined by binding six or more EA confined to the cell half-perimeter. Caps formed in serum were mainly extreme caps in which bound EA appeared as a morula contiguous with the adherent cell. At 21°C and 37°C in serum 80% live rosettes formed caps; virtually none formed at 4°C and about 25% were seen in PBS at 21°C. Similarly, 10% extreme caps in PBS and 60 and 70% in serum were seen at room temperature and 37°C, respectively, suggesting a serum factor(s) was important in promoting live rosette capping. Capping was reversibly inhibited by sodium azide although inhibition was incomplete even at 0.1 M, a concentration 10-fold higher than that giving complete inhibition of lymphocyte antigen-receptor capping. Azide also induced reversion of capped rosettes to diffuse, non-capped rosettes, and to levels comparable to those seen in inhibition studies. Re-exposure to EA after rosette capping failed to increase either the proportion of cells forming rosettes or the fullness of such rosettes indicating a critical number of receptors had been capped. Live rosettes induced a spherocytosis in bound EA irrespective of capping status; such changes appeared early and in PBS where capping was minimal. Dead cells bore EA as normal biconcave discs. Some rosetting EA were ultimately hemolyzed upon prolonged incubation, and erythrophagocytosis was seen in about 15% of capped rosettes at 37°C.
The Function of Macrophages in Antigen Recognition by Guinea Pig T Lymphocytes
Authors: Shevach Ethan M.
Published: December 2022

Activation of immune T lymphocytes by antigen-pulsed macrophages is mediated by the Ia antigens of the guinea pig MHC or the products of closely linked genes. Studies using combinations of macrophages and T cells derived from outbred animals with different Ia antigens and/or Ir gene products have demonstrated that sharing of Ir gene products between macrophage and T cells is not sufficient for effective macrophage-T cell interaction. The role of the Ia antigens in the absence of the linked Ir gene products could not be directly examined because we were unable to identify an animal which bore the full complement of Ia antigens in the absence of the Ir gene that is normally associated with them. The results of these studies support the concept of the functional expression of the Ir gene product in the macrophage.
Studies on T Cell Clonal Expansion
Authors: Kamat Ramesh , Henney Christopher S.
Published: December 2022

Spleen cells from C57BL/6 mice immunized with a DBA/2 mastocytoma (P815) were harvested at various stages of the immune response and cultured in vitro in the presence and absence of antigen.

Killer T cell activity in immune spleens could not be demonstrated until 6 or 7 days after antigen, but spleen cells harvested as early as 3 or 4 days and cultured for 24 hr at 37°C showed significant cytotoxicity. This increased activity was not augmented further by culturing with antigen.

“Memory” T cells, whose in vitro differentiation into killer cells required the presence of antigen, could not be demonstrated until 9 or 10 days after alloantigenic stimulation. Once produced, however, these cells persisted for at least 6 months. Memory cells, like killer T cells bound avidly to homologous allogeneic monolayers.

There were indications that the memory T cell pool was heterogeneous. On one hand, when cells harvested 10 days after stimulation were exposed to antigen in vitro, their lytic activity increased within 24 hr but showed no further increases when the culture period was extended. In contrast, 45-day-old immune cells showed increasing lytic activity throughout a 4-day exposure to antigen. Augmentation of lytic activity in both cell populations was independent of DNA synthesis through the first 24 hr of culture. Subsequent increases in the activity of 45-day cells was dependent upon cell proliferation.

Both the antigen-independent augmentation of lytic activity which followed culturing of immune cells, and the antigen-induced differentiation of memory cells were reversibly inhibited by a series of drugs which raised lymphocyte cAMP levels.
Preservation of a T Cell-Mediated Immune Response in Marek's Disease Virus-Infected Chickens by Vaccination with a Related Virus
Authors: Schierman Louis W. , Theis Gail A. , McBride Raymond A.
Published: December 2022

The oncogenic effect of Merek's disease virus (MDV)3 is manifested by a lymphoproliferative neoplastic condition primarily affecting nerves and visceral organs (For reviews see References 1, 2). Necrosis and atrophy of both the thymus and bursa of Fabricius are frequent findings in MDV-infected birds and degenerative changes are found in feather follicle epithelium where complete enveloped virions are formed. The necrobiosis in the thymus and bursa is associated with the presence of viral antigens but enveloped virions are very rare in these organs. There is evidence that the lymphoid tumors are composed of thymus-derived cells (3, 4) which possess tumor-associated antigens (5). Suppression of both cellular (6) and humoral (7) immune responses in MDV-infected chickens has been reported. In this regard, we have shown that spleen cells from birds infected with MDV are markedly depressed in their responsiveness to the T cell mitogen phytohemagglutinin (PHA) (8).
Protection against Experimental Allergic Encephalomyelitis with Peptides Derived from Myelin Basic Protein: Presence of Intact Encephalitogenic Site Is Essential
Authors: Driscoll Bernard F. , Kies Marian W. , Alvord Ellsworth C.
Published: December 2022

Experimental allergic encephalomyelitis (EAE) is a cell-mediated autoimmune response directed toward a component of central nervous system (CNS) tissue, myelin basic protein (BP). Injection of animals with either whole CNS tissue or purified BP in complete Freund's adjuvant (CFA) induces severe and usually fatal disease. Preimmunization of animals with BP in incomplete Freund's adjuvant (IFA) prevents EAE. We have examined the relative abilities of whole guinea pig BP and its fragments to protect guinea pigs from subsequent EAE induction. The data suggest that the presence of the intact encephalitogenic site (residues 113–121) in the molecules used for preimmunization is necessary but may not be sufficient for complete protection against EAE induction.
Immunosuppression of Experimental Autoimmune Myasthenia Gravis by Hydrocortisone and Azathioprine
Authors: Abramsky Oded , Tarrab-Hazdai Rebeca , Aharonov Aharon , Fuchs Sara
Published: December 2022

Experimental autoimmune myasthenia gravis (EAMG), induced in rabbits by injection of acetylcholine receptor (AChR) from Torpedo californica, was suppressed by appropriate treatment with hydrocortisone or with azathioprine. Administration of hydrocortisone in gradually increasing doses, starting at the time of immunization with the receptor, prevented exacerbation of the disease in the early stages of treatment, as was the case when hydrocortisone was administered in high doses from the beginning. Prolonged administration of the antimetabolite azathioprine (Imuran) prevented the appearance of EAMG, for at least 4 months, in rabbits immunized with AChR. Cell-mediated immunity to AChR was demonstrated to be significantly decreased in such treated animals.

The effects of hydrocortisone and azathioprine on EAMG support the view that the disease involves an immunologically cell-mediated mechanism and indicate that the experimental disease can serve as a useful model for chemotherapy of the human disease.
Eosinophil Chemotactic Factor (ECF)
Authors: Czarnetzki Beate M. , König Wolfgang , Lichtenstein Lawrence M.
Published: December 2022

Eosinophil chemotactic factor (ECF), previously thought to be primarily associated with human basophils and mast cells, could be released from human neutrophils (PMN) and eosinophils but not lymphocytes by the calcium ionophore A23187. Release of ECF from PMN was time and dose-dependent. Like antigen-induced, basophil-derived ECF, PMN-derived ECF had a high selectivity for eosinophils as determined by differential counts of migrating cells and by deactivation studies. Chromatographic analysis of PMN-derived ECF on Sephadex G-25 showed an elution pattern very similar to that of basophil-derived ECF. With rat peritoneal cells, it was possible to show that mast cells as well as mast cell-depleted cell preparations could be induced to release an ECF that appears to be similar or identical to human ECF. These findings suggest that ECF may play a role in inflammatory processes involving cells other than basophils and mast cells.
Blocking of MLC Stimulation by Anti-Ia Sera: Studies with the Virus Plaque Assay
Authors: Kano Shogo , Bloom Barry R. , Shreffler Donald C.
Published: December 2022

The development of congenic mouse strains identical at the H-2K and H-2D loci but differing by I-region associated (Ia) determinants has permitted an association to be established between Ia determinants and stimulation in mixed lymphocyte culture reactions (MLR). The present experiments were undertaken to establish whether the Ir-coded control of MLR operated at the level of recognition or of stimulation. Reciprocal MLR were established between A.TH and A.TL mouse spleen cells in the presence or absence of anti-Ia sera directed either at determinants of the stimulating or responding cells. The number of T cells responding was assessed by the virus plaque assay.

Anti-Ia sera directed against the responding cells were no more inhibitory of the MLR than normal mouse serum. In contrast, anti-Ia sera directed against determinants of the mitomycin-treated stimulating cells markedly inhibited activation of T cells in the MLR.
Analysis of Immunoglobulin Genes: DNA/RNA Hybridization with Immunoglobulin κ-Chain mRNA and Isolation and Translation of Hybridized RNA
Authors: Storb Ursula , Marvin Stephanie
Published: December 2022

Immunoglobulin κ-chain mRNA was hybridized with DNA in order to assess the κ-gene frequency. κ-mRNA was purified from membrane-bound ribosomes of mouse myeloma MOPC-41 by poly (U) chromatography and isolation of a 13S RNA by successive sucrose density gradient centrifugations. The RNA coded for κ-chain precursor molecules in cell-free protein synthesis and essentially no other proteins. MOPC-41 κ-mRNA hybridized with MOPC-41, MPC-11, and Krebs DNA with the same kinetics: the majority of the hybrids was formed with rare or unique DNA sequences (Cot/2 450 to 900), a small portion with highly repetitive sequences (Cot/2 5–6). The slow hybrids were well matched and the rapid hybrids were mismatched by about 4%, regardless of the DNA used.

It was further investigated whether the rapid hybrids contained translatable κ-mRNA or were due to impurities in the RNA preparations. κ-mRNA and globin-mRNA (as an internal standard for a unique transcript) were hybridized with DNA to Cot 20 or 48, the hybridized and unhybridized RNA were isolated by hydroxyopatite-urea chromatography and, after removal of the DNA, translated in a cell-free system. The cell-free products were analyzed by SDS-polyacrylamide gel electrophoresis and immunoprecipitation. It was found that approximately equal quantities of translatable κ- and globin-mRNA were hybridized maximally 1.7%). The results do not support the hypothesis that κ-mRNA is a transcript of both repetitive and unique DNA sequences.
Characterization of an Antiserum Specific for Cell Surface Antigens of Rat Mast Cells
Authors: Yiu Suk Hing , Froese Arnold
Published: December 2022

Rabbits were immunized with rat peritoneal mast cells (RMC) in complete Freund's adjuvant. The antisera (anti-RMC) were checked for their reactivity with RMC by intradermal skin tests in rats. The best serum was selected and absorbed with rat liver cells and rat immunoglobulins, including IgE. The absorbed serum (anti-RMCabs), as well as the anti-RMC serum, were then tested for their reactivity with RMC. Both sera were cytotoxic to RMC but only anti-RMC was cytotoxic for rat lymph node cells. Both sera gave positive reactions in rat skin, as seen by the permeability to Evan's blue dye. The binding of rat IgE to RMC was also inhibited by both sera. A control rabbit anti-rat sarcoma serum absorbed with liver cells did not show any interaction with RMC. When 125I-labeled RMC surface antigens were precipitated with anti-RMCabs and analyzed by SDS polyacrylamide gel electrophoresis, several components were observed. Among these was one with a mobility identical to that of a mast cell surface component that had previously been identified as the receptor for IgE or at least a component thereof.
The Requirement for Esterase Activation in the Anti-Immunoglobulin-Triggered Movement of B Lymphocytes
Authors: Becker Elmer L. , Unanue Emil R.
Published: December 2022

In this paper we bring evidence suggesting that there is activation of an esterase upon reaction of anti-immunoglobulin antibodies (anti-Ig) with murine B lymphocytes. B lymphocytes upon exposure to anti-Ig cap the ligand-receptor complexes and immediately afterward become briefly motile. It is this latter step which is inhibitable by exposure to di-isopropyl phosphofluoridate (DFP). Various experimental manipulations indicated that treatment with anti-Ig activates the cell for motility which, however, is not manifested until the temperature is raised to 37°C. The cell incubated with anti-Ig at cold temperatures becomes susceptible to the effect of DFP, suggesting that the antibody-treated cells are activated up to but not beyond the DFP inhibitable step. Exposure of cells to DFP and removal of it before their treatment with anti-Ig does not affect the anti-Ig-induced response. Four lines of evidence indicate that the reduction of lymphocyte movement by DFP is due to the inhibition of an esterase activated by the combination of antibody and cell: 1) The inhibition by DFP is irreversible; once DFP has reacted it can be washed away and the antibody-treated cell is still inhibited. 2) The inhibition increases with time of contact of lymphocytes and DFP and with the concentration of DFP. 3) A very poorly phosphorylating phosphonate, phenyl ethyl pentylphosphonate is completely inactive under conditions where an excellent phosphorylating phosphonate, p nitrophenyl ethyl pentyl-phosphonate maximally inactivated the cells' movement. 4) The amino acid esters, tosyl L arginine methyl ester and benzoyl arginine methyl ester specifically prevent the inactivation by DFP. The last finding suggests that tosyl L arginine methyl ester and benzoyl L arginine methyl ester might be substrates for the putative antibody-induced lymphocyte esterase.

Lymphocytes incubated with antibody in the cold for more than 30 min lose their ability to move when the temperature is raised, suggesting that there is a time-dependent deactivation of the cell.
Visualization of Type I and II Collagens in Tissue Sections by Immunohistologic Techniques
Authors: Wick Georg , Nowack Hans , Hahn Eckhart , Timpl Rupert , Miller Edward J.
Published: December 2022

Rabbit and rat antibodies were prepared against Type I and II collagens derived from bovine skin and articular cartilage, respectively. As judged by passive hemagglutination and radioimmune assays, these antibodies could be rendered generally specific for the type of collagen used for immunization by immunoadsorption. Thus, antibodies to Type II collagen did not cross-react with Type I and III collagens from skin. However, antibodies to Type I collagen still showed some cross-reaction with Type III collagen. Antibodies to Type I procollagen showed a negligible degree of cross-reaction with Type III procollagen.

These purified antibodies reacted strongly with bovine and human tissue collagen as demonstrated by indirect immunoflourescence. Antibodies to Type I collagen stained dermal tissue, perichondral tissue, kidney stroma, aortic tissue, and annulus fibrosus. Antibodies to Type II collagen stained mainly the hyaline matrix of rib cartilage and nucleus pulposus. The staining patterns with anti-Type I procollagen were similar but not identical to that found with antibodies to Type I collagen. Neither of these antibodies reacted with kidney glomerular basement membrane. These antibody reagents are recommended as a sensitive and rapid screening tool for studying tissue distribution of collagen under normal and pathologic conditions.
Divergent Effects of Cyclophosphamide Administration on Mononuclear Killer Cells: Quantitative Depletion of Cell Numbers Versus Qualitative Suppression of Functional Capabilities
Authors: Hunninghake Gary W. , Fauci Anthony S.
Published: December 2022

The effects of various regimens of cyclophosphamide administration on guinea pig peripheral blood leukocytes were studied. Cyclophosphamide-induced immunosuppression was assessed by the effect of drug administration on the proportions and absolute numbers of leukocyte populations, and by the effect on functional capabilities of unfractionated and adherent cell-depleted mononuclear cell suspensions as measured by the PHA-induced cellular cytotoxicity and antibody-dependent cellular cytotoxicity assays against chicken erythrocyte targets. Intraperitoneal administration of five daily doses of cyclophosphamide (5 mg/kg) caused a modest absolute leukopenia but no change in cytotoxic effector function of the mononuclear cells remaining in the circulation. As the dosage of cyclophosphamide was increased to 20 mg/kg/day to produce a pronounced leukopenia, a profound neutropenia (< 300 polymorphonuclear leukocytes/mm3) together with a marked decrease in mononuclear cell effector function was noted. A single i.p. injection of cyclophosphamide (100 mg/kg), which produced identical degrees of leukopenia of each leukocyte class as did daily administration of cyclophosphamide (20 mg/kg/day), caused no change in mononuclear cell effector function when compared to saline controls. Complement receptor-bearing and Fc-receptor bearing mononuclear cells were decreased to the same degree by both regimens of cyclophosphamide administration. Removal of adherent cells from mononuclear cell suspensions by column purification resulted in a marked decrease in cytotoxic effector function at low effector to target ratios. At higher effector to target ratios there was no difference in cytotoxic effector function between unfractionated and column-purified cells. In contrast, the functional defect in mononuclear cell suspensions from animals that received five daily doses of cyclophosphamide (20 mg/kg) could not be compensated for at higher effector to target ratios, indicating that this functional defect was not an artifact of relative depletion of monocytes by cyclophosphamide, but was due to an actual suppression of the effector functional capabilities of the killer cells. This study indicates that, dependent on the particular regimen of drug administration, the quantitative depletion of mononuclear cell populations by cyclophosphamide administration can be clearly distinguished from the qualitative effect on certain functional capabilities of surviving cells.
The Course of Schistosoma Mansoni Infection in Thymectomized Rats
Authors: Cioli Donato , Dennert Gunther
Published: December 2022

Inbred rats were thymectomized, irradiated, and reconstituted with T cell-free bone marrow cells. Thymectomized-reconstituted (B rats) and control rats were infected with Schistosoma mansoni cercariae and the number of worms recovered was determined at various times after infection. The extent of immunosuppression was assessed by two criteria: 1) response to an injection of sheep erythrocytes (plaque assay, hemagglutination, hemolysis); 2) response to schistosome antigens (passive hemagglutination). Humoral responses to worm antigens were completely suppressed in almost all instances and anti-sheep erythrocyte responses showed a more variable but always very definite depression in B rats. The number of worms in B rats was about 4 times higher than in control animals at 5 weeks and about 3 times higher at 6 weeks. In a different experiment, rats were perfused at 4, 6, and 9 weeks after infection and the number of worms was found to be consistently higher in B rats, by a factor of about 2 at 4 weeks to a factor of about 4 or 6 at subsequent times. Although B rats had more worms than controls even at 9 weeks, a slow drop in their worm burden was noticeable with time in both experiments. Moreover, the size of worms in B rats was smaller than in controls and even 9-week-old worms failed to develop to normal size and appearance and could not be shown to produce fertile eggs.

These experiments show a definite involvement of the immune system in the “self-cure” phenomenon, but may at the same time suggest that other non-immune mechanisms are involved in determining the pattern of S. mansoni infection in the rat.
Structural Studies on Induced Antibodies with Defined Idiotypic Specificities
Authors: Capra J. Donald , Berek Claudia , Eichmann Klaus
Published: December 2022

The murine VH genetic map consists of two different subloci of VH genes. Seven idiotypes have been allocated to these two different subloci, one of which contains genes coding for the A5A idiotype, while the second contains the genes encoding for six other idiotypic markers. Previous studies from this laboratory have shown that anti-Ars antibodies are best classified in the murine VHII subgroup. Studies from other laboratories indicate that the J558 idiotype could be classified in a similar fashion. The sequence of the T15 myeloma protein VH region is most characteristic of the murine VHIII subgroup. These studies were undertaken in order to study the VH subgroup of two additional murine VH markers. Since one of these markers has been allocated to a separate sublocus, comparisons can be made for the first time between the “framework” residues (VH subgroup) and the murine genetic map. The data indicate that within a single presently defined sublocus proteins exist which contain remarkably different “framework” residues.
Isolation and Preliminary Characterization of Two Varieties of Low Molecular Weight Immunoglobulin in the Bullfrog, Rana Catesbeiana
Authors: Green Christopher , Steiner Lisa A.
Published: December 2022

Two varieties of low m.w. immunoglobulins have been isolated from the serum of Rana catesbeiana frogs. They are highly cross-reactive, although each also contains unique antigenic determinants. Since both low m.w. immunoglobulins were identified in the serum of 22 individual frogs, it was concluded that they are isotypic variants. The light chains of R. catesbeiana and mammalian high and low m.w. immunoglobulins are similar in electrophoretic mobility on polyacrylamide gels containing sodium dodecyl sulfate. The heavy chains of frog high m.w. immunoglobulins have the mobility of mammalian µ-chains; the heavy chains of both variants of frog low m.w. immunoglobulins migrate between mammalian µ- and γ-chains, in approximately the position of mammalian α-chains. An unusual structural feature of the R. catesbeiana high and low m.w. immunoglobulins is that the unreduced proteins are partially dissociated in sodium dodecyl sulfate.
Function of 2-Mercaptoethanol as a Macrophage Substitute in the Primary Immune Response in Vitro
Authors: Lemke Hilmar , Opitz Hans-Georg
Published: December 2022

The mechanism by which 2-ME acts as a macrophagesubstitute for the induction of a primary PFC response to SRC in vitro was studied in macrophage-depleted mouse spleen cell cultures. 2-ME could replace macrophages only in FCS-supplemented cultures. Evidence is presented that the function of 2-ME is independent of residual macrophages. Neither normal nor macrophage-depleted spleen cell cultures from congenitally athymic nude mice supplemented with 2-ME, with or without FCS, could give rise to a primary in vitro anti-SRC immune response.

2-ME, at an optimal concentration of 10-5 M, induced DNA synthesis in normal and macrophage-depleted spleen cells in both FCS-containing and serum-free cultures. The peak response occurred on day 3. This stimulation was accompanied by a polyclonal B cell activation to antibody secretion which was much more pronounced in FCS-containing than in serum-free cultures. Spleen cells from nude mice showed a weaker DNA stimulation than did cells from normal mice in FCS-containing cultures, and nearly no response under serum-free conditions. T cells obtained by a nylon column adherence method from normal mouse spleen cells showed good DNA synthetic responses in FCS-containing, but no response in serum-free cultures.

These results show that 2-ME has weak mitogenic activity for B cells, and in combination with FCS, strong mitogenic activity for T cells. Since the macrophage provides stimulation to the T cell in the primary anti-SRC PFC response in vitro, these results suggest that the direct mitogenic activity of 2-ME with FCS on T cells provides the functional substitution for macrophages.
Purification of Native Properdin by Reversed Affinity Chromatography and Its Activation by Proteolytic Enzymes
Authors: Minta Joe O.
Published: December 2022

A highly purified preparation of human properdin (P) has been obtained in a simple two-step procedure utilizing reversed application of the technique of affinity chromatography. The method involved precipitation of properdin from human serum and subsequent passage through an immunoadsorbent column of Sepharose anti-RP globulin which bound the contaminating proteins. The immunochemical properties of the isolated properdin (P) was found to be different from those of activated properdin (P̄). P was shown to be a 6.1S, β2 globulin with a mean subunit m.w. of 57,900. P̄ on the other hand was a 5.1S protein with γ2 mobility and a subunit m.w. of 53,000 daltons. Double diffusion analysis using anti-P̄ revealed a reaction of identity between P and P̄. However, when the reaction was developed with anti-P, a reaction of partial identity was obtained and the precipitin line of P was seen to spur over the line developed with P̄. Mild treatment with plasmin or trypsin converted P to P̄.

Unlike P̄, P was ineffective in triggering the activation of the Properdin System in RP unless trace amounts of zymosan were added. Under these conditions P was found to be converted to P̄. The results indicate that properdin is present in fresh serum in a precursor form and its activation to P̄ involves a limited proteolytic cleavage of the molecule.
In Vitro Stimulation of Neutrophil Motility by Levamisole: Maintenance of cGMP Levels in Chemotactically Stimulated Levamisole-Treated Neutrophils
Authors: Anderson Ronald , Glover Annemarie , Koornhof Hendrik J. , Rabson Arthur R.
Published: December 2022

Levamisole at concentrations of 10-3 M or 10-4 M consistently increased neutrophil random motility and chemokinesis (stimulated random migration). Similar concentrations also increased directional movement of polymorphonuclear leukocytes to both endotoxin-activated serum and hydrolyzed casein. This effect on chemotaxis was due to a true stimulation and was not due solely to increased random movement. The effect of levamisole on the neutrophils could be removed by washing, but persisted if the cells were initially treated with levamisole and serum or endotoxinactivated serum. After neutrophil stimulation with chemotactic factor an initial rise in intracellular cyclic AMP levels was detected which was not influenced by prior levamisole treatment. Intracellular cyclic GMP levels after an initial slight depression, returned to resting levels and gradually diminished over a 60-minute period. Levamisole treated cells consistently showed higher cyclic GMP levels and it is postulated that by maintaining intracellular cyclic GMP levels, microtubular assembly and cell motility might be enhanced.
Lack of Association of Human “B Cell” Antigens and Fc Receptor Activity of Antibody-Dependent Cytotoxic Lymphocytes
Authors: Miller June L. , Humphreys Robert E. , Mann Dean L.
Published: December 2022

Heterologous antisera directed against human B cell antigens were tested for their ability to inhibit the function of Fc receptors on human lymphocytes. These antisera coated on the attacking cells in a lymphocyte-dependent antibody-mediated cytotoxic assay inhibited lysis of sensitized target cells.

However, the F(ab)2 fragments of the IgG fraction of an anti-human Ia antiserum failed to inhibit the killing of sensitized targets by attacking cells. Antisera against other lymphocyte membrane antigens also inhibited target cell lysis when coated on attacking cells. These results suggest that there is not identity between human B cell antigens and Fc receptors of antibody-dependent cytotoxic lymphocytes.
Rat T Lymphocyte-Specific Antigens and Their Cross-Reactivity with Mouse T Cells
Authors: Ishii Yoshifumi , Koshiba Hirofumi , Yamaoka Hiroshi , Kikuchi Kokichi
Published: December 2022

Anti-rat T lymphocyte serum (ATLS)2 was prepared by immunizing rabbits with purified T cells from rat mesenteric nodes and absorbed with rat red cells and syngeneic sarcoma cells. The specificity of ATLS for rat T cells was confirmed by the following reasons: a) ATLS was not toxic for bone marrow cells but lysed most of the thymocytes and a number of spleen and lymph node cells, which were inversely correlated to the percentage of cells with B cell characteristics in respective organs; b) anatomical localization of ATLS-reactive cells in lymphoid organs coincided to the thymus-dependent areas, i.e., the paracortex of lymph node and the periarteriolar region of spleen; c) spleen cells treated with ATLS and complement failed to respond to phytohemagglutinin but normally responded to bacterial lipopolysaccharide; d) those cells treated with ATLS and complement could not induce a graft-vs-host reaction in F1 hosts, whereas the same treatment did not affect direct plaque-forming cells. All of these data confirm the specificity of ATLS and indicate that ATLS recognizes rat T lymphocyte-specific antigens (RTLA). Absorption studies showed that RTLA were present in higher concentration on medullary thymocytes and peripheral T cells than on cortical thymocytes, but absent from bone marrow, liver, and brain tissues. When the cross-reactivity of RTLA with mouse T cells was studied by C-dependent cytotoxicity and immunofluorescence, it was found that mouse T cells shared at least one determinant of RTLA with rat T cells, and that distribution pattern of the cross-reacting antigens in mouse lymphoid tissues was essentially the same as that of RTLA in rat lymphoid organs.
Generation of Cytolytic T Lymphocytes in Vitro
Authors: Fitch Frank W. , Ramseier Hansruedy
Published: December 2022

Antibody reactive with “recognition structures” (RS) of mouse lymphoid cells for alloantigens (anti-RS) was prepared by immunization of F1 hybrid mice with parental strain lymphoid cells or with antibody produced in one parental strain against alloantigens of the other parental strain. Such antisera prevented generation of the “product of antigenic recognition” (PAR) that is produced within a few hours in cultures prepared with a mixture of lymphoid cells from genetically disparate mice. However, treatment of responding lymphoid cells with anti-RS sera and complement did not inhibit generation of cytolytic T lymphocytes (CTL) in mixed lymphocyte cultures (MLC). Treatment of cells obtained from MLC with anti-RS sera and complement failed to inhibit cytolytic activity of such cells for specific alloantigens.
T Lymphocyte-Enriched Murine Peritoneal Exudate Cells
Authors: Schwartz Ronald H. , David Chella S. , Sachs David H. , Paul William E.
Published: December 2022

The recent development of a reliable murine T lymphocyte proliferation assay has facilitated the study of T lymphocyte function in vitro. In this paper, the effect of anti-histocompatibility antisera on the proliferative response was investigated. The continuous presence of anti-Ia antisera in the cultures was found to inhibit the responses to the antigens poly (Glu58Lys38Tyr4) [GLT], poly (Tyr, Glu) poly D,L Ala—poly Lys [(T,G)-A—L], poly (Phe, Glu)-poly D,L Ala—poly Lys [(ø,G)-A—L], lactate dehydrogenase H4, staphylococcal nuclease, and the IgA myeloma protein, TEPC 15. The T lymphocyte proliferative responses to all of these antigens have previously been shown to be under the genetic control of major histocompatibilitylinked immune response genes. The anti-Ia antisera were also capable of inhibiting proliferative responses to antigens such as PPD, to which all strains respond. In contrast, antisera directed solely against H-2K or H-2D antigens did not give significant inhibition. Anti-Ia antisera capable of reacting with antigens coded for by genetically defined subregions of the I locus were capable of completely inhibiting the proliferative response. In the two cases studied, GLT and (T,G)-A—L, an Ir gene controlling the T lymphocyte proliferative response to the antigen had been previously mapped to the same subregion as that which coded for the Ia antigens recognized by the blocking antisera. Finally, in F1 hybrids between responder and nonresponder strains, the anti-Ia antisera showed haplotype-specific inhibition. That is, anti-Ia antisera directed against the responder haplotype could completely block the antigen response controlled by Ir genes of that haplotype; anti-Ia antisera directed against Ia antigens of the nonresponder haplotype gave only partial or no inhibition. Since this selective inhibition was reciprocal depending on which antigen was used, it suggested that the mechanism of anti-Ia antisera inhibition was not cell killing or a nonspecific turning off of the cell but rather a blockade of antigen stimulation at the cell surface. Furthermore, the selective inhibition demonstrates a phenotypic linkage between Ir gene products and Ia antigens at the cell surface. These results, coupled with the known genetic linkage of Ir genes and the genes coding for Ia antigens, suggest that Ia antigens are determinants on Ir gene products.
Murine Plasma Cells Secreting More Than One Class of Immunoglobulin Heavy Chain
Authors: Morse Herbert C. , Pumphrey Janet G. , Potter Michael , Asofsky Richard
Published: December 2022

Primary ascites from 778 consecutive plasmacytomas of BALB/c and other mouse strains were tested by immunoelectrophoresis for the presence of paraproteins belonging to different immunoglobulin classes. γA was produced by 338 tumors; γGs (γ1, γ2a, γ2b, or γ3) by 163 tumors; and γM by seven tumors. Free κ- or λ-type light chains were produced by 14 tumors, and no detectable paraprotein was produced by 202 tumors. Two or more paraproteins were detected in the primary ascites of 54 plasmacytomas. Evidence suggests that in most cases, the multiple paraproteins are produced by simultaneously occurring, independent plasma cell tumors. However, two tumors, SAMM 368 and TEPC 609, produced two paraproteins for over 30 generations in transplantation. This finding implies that single clones of plasma cells have the capacity to produce more than one class of immunoglobulin.
Analysis of the Diversity of Murine Antibodies to Dextran B1355
Authors: Hansburg Daniel , Briles David E. , Davie Joseph M.
Published: December 2022

Mice immunized with a combination of dextran B1355 in adjuvant followed by three injections of 2 × 109 Escherichia coli B organisms produced an average of 14.5 mg/ml of anti-dextran antibodies. It was demonstrated that the stimulating effect of E. coli B was due to antigenic determinants cross-reactive with B1355 and not solely because of adjuvant properties of the organism. The anti-dextran antibodies were distributed among both 7S and 19S components. Isoelectric focusing of the 7S antibodies showed several spectrotypes of antibody, most of which were shared by the majority of the individual sera. The limited spectrotypic heterogeneity of the 7S antibodies was supported by idiotypic studies. Thus, a heterologous, anti-idiotypic serum, rabbit anti-M104, was prepared which distinguished between two closely related myeloma proteins, M104 and J558, with specificity for α-(1 → 3) dextran. This antiserum demonstrated that some, but not all, of the 7S and 19S anti-dextran antibodies possessed variable region determinants cross-reactive with M104.