Horse Antiserum Research Articles

A reagin-like antibody in horse serum. II. Anti-human IgE induced reversed cutaneous anaphylaxis-like responses in horse skin.

Fc specific anti-human IgE serum induced prolonged reversed cutaneous anaphylaxis (RCA)-like reactions in horse skin. Morphologically and histologically, these reactions resembled passively induced late cutaneous anaphylaxis responses in human skin, but differed from reversed passive Arthus responses induced in horse skin using anti-horse IgG serum. The induction of RCA-like responses in horse skin by anti-human IgE indicates shared Fc antigenic determinants on human IgE and a horse homocytotropic or reagin-like antibody.

Neurotoxin-specific immunoglobulins accelerate dissociation of the neurotoxin-acetylcholine receptor complex.

Toxin isolated from cobra venom and labeled with tritium was incubated with membranes rich in acetylcholine receptors. The amount of toxin bound to the receptors was determined and the kinetics of dissociation of the receptor-toxin complex was followed. Addition of an excess of horse antiserum to the venom resulted in a significant acceleration of the dissociation reaction. Similarly, a monoclonal antibody against the toxin accelerated dissociation of the receptor-toxin complex. The results indicate that specific antibody binding destabilizes the toxin-receptor complex.

Titration of antiserum to South American rattlesnake ( Crotalus durissus terrificus) venom by inhibition of phospholipase A 2 activity

Horse antiserum to the venom of Crotalus durissus terrificus, A South American rattlesnake, inhibits the phospholipase activity of the crude venom. There is a close relationship between this inhibitory property and the neutralizing potency of the antiserum in vivo. This may provide the basis for a rigorous standardization of anticrotalid venom in vitro.

Human T lymphocytes of inducer and suppressor type occupy different microenvironments.

Distinct subsets of human peripheral T-cell populations have recently been characterized using mouse monoclonal antibodies and conventional hetero-antisera in functional tests in vitro1–6. ‘Inducer’ or ‘helper’ T cells have been shown to react with a monoclonal antibody termed OKT4. These OKT4+ cells respond to soluble antigens3, help B-lymphocyte differentiation into plasma cells in pokeweed mitogen stimulated cultures4 and assist the development of cytotoxic T cells in mixed lymphocyte cultures (MLC)3. In contrast, the ‘suppressor-cytotoxic’ T-cell subset is recognized by the monoclonal antibodies OKT55 and OKT86. The same subset can also be labelled with a conventional horse antiserum which (after extensive absorption) recognizes TH2 antigen5,7. These OKT8+, TH+2 cells fail to respond optimally to soluble antigen3 but contain the concanavalin-A induced suppressor cell population5,7 and the cells which develop cytotoxic activity in mixed lymphocyte reaction (MLR)-induced cell-mediated lympholysis3. The physiological role, tissue distribution and recirculation patterns of ‘inducer’ and ‘suppressor’ T cells are unknown, although changes in the proportion and activity of blood-borne T-ceU subsets have been observed in various immunoregulatory disorders8,9. We have therefore now analysed the distribution of these T-cell subsets in the lymphohaematopoietic organs and the gut. OKT4+, OKT8− cells of inducer type predominate in the thymic medulla, blood and T-cell traffic areas such as tonsillar paracortex and intestinal lamina propria. OKT4−, OKT8+; cells of suppressor-cytotoxic type, on the other hand, constitute the larger part of T-cell population in normal human bone marrow and gut epithelium. Furthermore, a close micro-anatomical relation can be seen between the OKT4+, OKT8−T cells and non-lymphoid cells expressing large amounts of la-like (p 28,33) antigens, for example, interdigitating (ID) cells and Ia+ macrophages, suggesting that Ia-like antigens may play a part in the local regulation of inducer T cell activity. Thus the T-cell subsets which have been shown to have different functions in vitro seem also to have different patterns of tissue distribution, implying different immunological functions in vivo.

Improved antiserum agar method for the serogroup differentiation of Neisseria meningitidis Y and W135.

Rabbit antisera prepared to meningococcal serogroups Y and W135 strains were compared with horse antisera using the antiserum agar method (ASA) for the serogroup identification of Neisseria meningitidis. Thirty-seven group Y strains formed immunoprecipitates with the Y rabbit serum only, whereas the same Y strains formed immunoprecipitates with both the Y and W135 horse antisera. Forty-seven W135 strains formed specific immunoprecipitates with both the rabbit and horse W135 antisera by ASA. None of the 166 meningococcal isolates, representative of other meningococcal serogroups, formed immunoprecipitates with the groups Y and W135 rabbit or horse antisera. Use of specific Y and W135 rabbit antisera in ASA provides an improved technique for the serogroup differentiation of groups Y and W135 meningococci.

Suppressor T cells in tolerance to deaggregated horse anti-human thymocyte globulin in man.

To understand the mechanism by which deaggregated horse anti-human thymocyte globulin (dATG) fails to induce untoward immunological reactions in man, three patients who received ATG and two patients who received dATG were studied for evidence of sensitization or tolerance to the foreign globulin. The ATG but not the dATG recipients developed allergic or serum sickness reactions; antihorse serum antibody could be detected in their serum and their blood cells proliferated in vitro in the presence of horse serum and secreted antihorse serum antibodies (P less than 0.001). Tolerance of the dATG recipients was shown to be mediated by specific T suppressor cells that carry receptors for horse serum and could be detected in the blood of one patient, whom we have studied serially, 13 weeks after therapy. Deaggregated ATG does not induce allergic or serum sickness reactions. It induces tolerance to the foreign protein via the generation of specific short-lived suppressor cells.

Generalized viral illness caused by an intermediate strain of adenovirus (21/H21 + 35).

Ten isolates of an antigenically intermediate strain of adenovirus were recovered from eight persons who presented febrile upper respiratory illness, pneumonia, or multisystem disease between November 1976 and August 1978. Six isolates were recovered from four members of an extended family in which the other nine members all had serologic evidence of infection; the other four isolates were epidemiologically unrelated. The isolates were placed in hemagglutination group IB on the basis of results of differential hemagglutination tests. All isolates were identified as adenovirus 21 by serum-neutralization tests and as types 21, 34, or 35 by hemagglutination-inhibition tests with reference rabbit and horse antisera. Conversely, rabbit antiserum to the intermediate strain had high titers of neutralizing antibody to adenovirus 21 and high titers of hemagglutination-inhibiting antibody to adenoviruses 21 and 35. The patients' serologic responses were similar. Thus, the isolates were typed as adenovirus 21/H21 + 35 and are the first such intermediate strains recorded.

A rapid screening method for the detection of monospecific antibodies against hemoglobins

A simple procedure for the detection of monospecific antibody against C57BL/6 mouse hemoglobin that would not cross-react with DBA/2 mouse hemoglobin is described. The horse antiserum against C57BL/6 mouse hemoglobin is absorbed with DBA/2 mouse hemoglobin. The absorbed serum is then allowed to react with small amounts of DBA/2 hemoglobin-Sepharose and C57BL/6 hemoglobin-Sepharose in separate tubes, followed by reaction with fluorescein isothiocyanate-conjugated goat antihorse IgG. A strong fluorescence in the C57BL/6 immunoabsorbent and little or no fluorescence in the DBA/2 immunoabsorbent show the presence of monospecific antibody against C57BL/6 hemoglobin. The method has general applicability as antibodies against other hemoglobins and proteins can be detected.

Localization in Rat Skin Transplants of Purified 125l-Labeled Xenogeneic Histocompatibility Antibody

It is shown that appropriate xenogeneic rabbit and horse antiserum, as well as allogeneic rat serum, can serve as sources from which /sup 125/I-labeled antibody to a strong histocompatibility antigen expressed on Fischer-344 strain rat cells can be prepared. After iv administration all three types of antibody would localize preferentially in F-344 skin grafts made 4 days earlier on Buffalo strain rats. After 24 hr, /sup 125/I localization ranged from 10 to 20 percent of the injected dose on F-344 skin grafts averaging 0.3 g weight on 150 to 190-g Buffalo rats. The horse serum used as an antibody source was a commercially available antirat-lymphocyte serum. Prior ip administration of two or three doses of 0.5 ml of this serum largely blocked uptake by F-344 skin grafts of both the /sup 125/I-labeled alloantibody and the /sup 125/I-labeled xenogeneic histocompatibility antibody prepared from this horse antiserum.

Application of pepsin-digested horse antibodies for quantitative immunoelectrophoresis

The majority of the precipitating antibodies in hyperimmune horse serum belong to the beta globulins, as demonstrated by reversed immunoelectrophoresis. As these antibodies migrate in agarose gel during electrophoresis in conventional pH = 8.6-8.9 buffers, horse antiserum cannot be used satisfactorily for quantitative immunoelectrophoresis. On pepsin digestion of horse antiserum F(ab)' 2 fragments are generated which migrate with the gamma globulins. This cleaved product works well in quantitative immunoelectrophoretic techniques.

Effect of pH and Haem Compounds on the Killing of Pasteurella septica by Specific Antiserum

The killing of Pasteurella septica by horse antiserum has features not previously associated with serum bactericidal reactions. The present work showed that lowering the pH from 7-4 to 6-8 abolished the action of antiserum. The bactericidal effect and the degradation of RNA seen when antiserum is added to P. septica growing in horse serum, were abolished at pH 6-8 in much the same way as when haem compounds were added to the system. Addition of chloramphenicol, rifampicin or puromycin to P. septica growing apparently normally in antiserum at pH 6-8 or in antiserum containing haem compounds led to rapid killing of the bacteria and to degration of their RNA. Addition of these antibiotics to P. septica growing in normal serum produced only bacteriostasis and did not induce RNA breakdown. In contrast, nalidixic acid, although inhibiting growth, did not induce rapid killing and RNA breakdown under the same conditions. These findings were unexpected and led to a reassessment of ideas concerning the mechanism of action of specific antiserum to P. septica. Although iron compounds clearly abolish the bactericidal based simply on an interference with bacterial iron supply is no longer sufficient. The process is more complex and must involve other factors.

Studies on complement-fixation reaction in equine infectious anemia. II. Identification of complement-fixing inhibitors.

The substances responsible for inhibiting complement-fixation (CF) reaction of the late-stage serum of an equine infectious anemia (EIA)-infected horse were investigated. It was found that the IgG and IgG(T) classes in the late-stage serum were responsible for the CF inhibition. IgA could not be detected in partially purified IgG(T) by an immunodiffusion test using rabbit anti-human IgA serum. Other serum components could not be demonstrated in purified IgG by immunoelectrophoresis using rabbit anti-horse serum. The IgG class simultaneously showed CF and CF-inhibiting (CFI) activities, whereas the IgG(T) class showed only CFI activity. The IgG(T) class could exert CFI activity only when it had been reacted with the EIA antigen before addition of the reference CF serum and complement. In contrast, the IgG class converted the CF-active reference serum into a non-CF-reactive one irrespective of whether it was simultaneously reacted with the EIA antigen and the reference CF serum, whether it was added to the reaction mixture of the EIA antigen and the reference CF serum, or whether it was sensitized with the EIA antigen before addition of the reference CF serum. Inhibitory activities of the IgG and IgG(T) classes seemed to be different from each other in their reaction pattern as far as tested under our experimental conditions. Their CFI activities seemed to be specific for EIA, being negative in CFI activity in reaction with other antigens.

Rapid degradation of ribosomal RNA in Pasteurella septica induced by specific antiserum

Abstract Antibody, transferrin and complement, acting together, directly interfere with the biochemistry of Pasteurella septica effecting an inhibition of RNA accumulation and the cessation of bacterial growth. This communication describes the fate of ribosomal RNA in antiserum inhibited cells. P. septica RNA was prelabelled with 32 P or [ 14 C] uracil, the organisms resuspended in unlabelled horse serum and antiserum added. The label remained in the RNA for up to 45 min after the addition of antiserum but was lost very rapidly thereafter. By the 90th min, 80–90% had been lost, this loss being reflected by the appearance of labelled material in the serum. Sucrose gradient analysis showed that the ribosomes were lost from such inhibited cells and that the ribosomal RNA was broken down before it was lost from the cells or even released from the ribosomal particles. Addition of haematin to the system prevented both the inhibition of bacterial growth and the subsequent breakdown of RNA. A similar degradation of ribosomal RNA was seen when antiserum was added to P. septica growing in rabbit serum, although in this case, antiserum produced bacteriostasis instead of the lethal effect seen in horse serum.

Thymus-dependent lymphocytes in tissue sections of rejecting rat renal allografts.

Lewis kidneys were grafted into BN recipients and examined at daily intervals up to 6 days after grafting with immunofluorescent reagents. A horse antiserum specific for T lymphocytes revealed an increasing number of T lymphocytes in the cellular infiltrates of rejecting allografts. These were detectable 1 day after grafting, reached a maximum 3 days later, and were relatively diminished at 6 days. In control isografts and nonimmunological inflammations of kidney, a small number of dispersed T lymphocytes was seen. A rabbit antirat thymocyte antiserum, given to allografted BN rats, prolonged survival of the grafts and decreased the cellular infiltrate and the number of T lymphocytes in the infiltrates. We conclude that in graft rejection there is a flow of T lymphocytes into areas of tissue damage and these T lymphocytes are immunologically reactive to graft antigens.

Mechanism of recovery from systemic infection with vaccinia virus. 3. Effects of antithymocyte serum.

Treatment with rabbit or horse antiserum to murine thymocytes (antithymocyte serum or ATS) potentiated primary systemic infection with vaccinia virus in mice. Formation of serum neutralizing antibody was suppressed and viremia was prolonged by treatment with ATS. This potentiation of vaccinia virus infection by ATS treatment was largely reversed by passive transfer of physiologic amounts of antibody late in the course of infection, at a time when nonimmunosuppressed mice had neutralizing antibody in their sera. The concentration of interferon in serum was not affected by treatment with ATS. These results suggest a critical role for antibody, but not for cellular immunity, in recovery of mice from primary systemic infection with vaccinia virus. The relative importance of cellular immunity, humoral antibody, and of nonimmune factors, such as interferon, in recovery from a primary viral infection has not been established. In recent years evidence from both infections in man and experimental studies has been interpreted to suggest a crucial role for cellular immunity and a less important role for humoral antibody in this recovery process, particularly for vaccinia virus infections [1-3]. To test this hypothesis, I have conducted a series of studies to determine the effect of a number of different immunosuppressive agents on primary vaccinia virus infection in mice. These studies have suggested that humoral antibody, but not cellular immunity, is critical in recovery from primary vaccinia virus infection in mice [4]. The present communication specifies the effect of horse and rabbit antisera to murine thymocytes (antithymocyte serum or ATS) on the course of primary vaccinia virus infection of the mouse.

Sjögren-Larsson syndrome. Report of two cases in twins.

<h3>Abstract</h3> COVID-19 caused by the emerging human coronavirus, SARS-CoV-2, has become a global pandemic, leading a serious threat to human health. So far, there is none vaccines or specific antiviral drugs approved for that. Therapeutic antibodies for SARS-CoV-2, was obtained from hyper immune equine plasma in this study. Herein, SARS-CoV-2 RBD with gram level were obtained through Chinese hamster ovary cells high-density fermentation. The binding of RBD to SARS-CoV-2 receptor, human ACE2, was verified and the efficacy of RBD in vivo was tested on mice and then on horses. As a result, RBD triggered high-titer neutralizing antibodies in vivo, and immunoglobulin fragment F(ab’)₂ was prepared from horse antisera through removing Fc. Neutralization test demonstrated that RBD-specific F(ab’)₂ inhibited SARS-CoV-2 with EC₅₀ at 0.07 μg/ml, showing a potent inhibitory effect on SARS-CoV-2. These results highlights as RBD-specific F(ab’)₂ as therapeutic candidate for SARS-CoV-2.

Evaluation of various antisera and gels for detection of hepatitis-associated antigen by immunodiffusion and immunoelectroosmophoresis tests.

Six antisera to hepatitis-associated antigen (HAA) were compared for sensitivity in detecting the antigen in gel diffusion and immunoelectroosmophoresis(IEOP) tests; these included three human antisera, two guinea pigantisera, and one horse antiserum. Gels and reagents from three commercial sources (Courtland, Hyland, and Spectra) and from this laboratory were compared for reliability in detecting HAA by gel diffusion and IEOP tests. In gel diffusion tests two human antisera, one distributed by the National Institutes of Health and one from this laboratory, were the most sensitive, detecting antigen in 83% of the sera positive by complement fixation (CF). Gels from Courtland Laboratories and this laboratory gave the greatest numbers of positive immunodiffusion reactions. The use of an enhancement pattern (testing each serum adjacent to a positive antigen) increased the sensitivity of the gel diffusion test for detecting HAA-positive sera from 57 to 83%. In IEOP tests, the three human antisera detected HAA in 91 to 95% of the sera positive by CF, and the animal antisera in only 83 to 84%. The importance of careful reading of tests was emphasized by the fact that the antiserum which gave the greatest number of positive reactions in both test systems showed the weakest lines of precipitate.

Occurrence and persistence of "Australia" antigen determinants.

"Australia" (Au) antigens from various groups of individuals were examined for the presence of d and y determinants. Antigens from all of the 214 individuals examined were found to possess either the d or y determinant, in addition to the a determinant. With the ay antiserum employed, antibody absorption was found to be a more effective means than demonstration of spur formation for detection of the y determinant. Antigens with ad specificity predominated in a collection of sera from non-ill Tongan children and adults, but no significant differences were noted in the specificities of antigens from individuals from four different regions. Almost all of the antigens from various groups of individuals in California, including inmates of a state hospital, a group of heroin users, and hepatitis patients from San Francisco General Hospital, were of the ay specificity. With one exception, antigenic specificities were found to persist for 3 years in a group of Tongan school children. Specificities also persisted in chronic carriers from California and in hepatitis patients over the course of antigenemia. Of 15 human and 4 animal antisera examined, antibodies to the y determinant were demonstrable only in a single (human) antiserum, and antibodies to the d determinant were demonstrable in one guinea pig antiserum and one horse antiserum.

The effect of pretreatment of allogeneic bone marrow graft recipients with antilymphocytic serum on the acute graft-versus-host reaction in monkeys.

SUMMARY Short-term pretreatment of irradiated, bone marrow-treated monkeys with horse antilymphocyte serum (ALS) roughly doubles the survival time by decreasing the severity and delaying the onset of acute secondary disease in random host-donor combinations. It was found that (1) horse anti monkey serum does not interfere with the take and proliferation of the graft, and that (2) rabbit anti monkey serum acts less selectively, as shown by a high incidence of transplant failures. The absence of signs of stem cell toxicity in monkeys treated with horse ALS cannot be attributed to a lower degree of immunosuppression, because lower doses of rabbit ALS are as toxic as larger doses. The use of ALS for pretreatment of human bone marrow recipients is recommended, but, for the time being, only horse serum should be used. The effective dose of horse ALS for monkeys is in the order of 60 mg of IgG/ kg body wt/ day. Pretreatment for 2 days seems to be adequate

Killing of mycoplasmas by the antibodies to foreign antigens acquired by the organisms from the growth medium.

The incubation of rabbit anti-horse serum (anti-HS) with mycoplasmas (represented byMycoplasma gallisepticum andMycoplasma pneumoniae strains) grown in presence of 20% horse serum resulted in significant killing of the test organisms. In simultaneous tests, however, the anti-HS failed to kill the same mycoplasma strains grown in presence of rabbit serum. The killing of the test mycoplasmas was abolished when the anti-HS was heated at 56°C for 30 min. Killing by anti-HS failed to occur in presence of M/100 EDTA. It is suggested that the killing of the mycoplasmas observed by the anti-HS is a complement mediated reaction, the antibody being specifically directed against the horse serum antigenic determinant(s) acquired by the mycoplasma cell membrane during theirin vitro cultivation in the presence of horse serum. The reaction observed is analogous to the phenomenon of “passive immune kill” reported in case of mammalian cells and “passive immune haemolysis” or “passive immune haemagglutination” in case of erythrocytes. The possible implications of this finding have been pointed out.