Infusion Of Antigen Research Articles

Immune response in the bovine mammary gland after intestinal, local, and systemic immunization.

The immune response in mammary glands of cattle was measured after intestinal, local, and systemic immunization with T4 bacteriophage. Nonlactating pregnant cows were immunized by infusions into the intestine or mammary gland and by subcutaneous injections in the region of the prescapular or external inguinal lymph nodes. Titers of antibodies of different isotypes were measured in serum and in lacteal secretions by enzyme-linked immunosorbent assay, and numbers of cells producing antibodies of each isotype were determined in lacteal secretions by the Jerne plaque assay. Substantial increases in immunoglobulin G subclass 1 (IgG1) and IgG2 antibody titers were detected in serum and lacteal secretions of animals immunized through an intestinal fistula. IgM and IgA antibody responses were low or undetectable. Low numbers of IgA and IgG1 plaque-forming cells were occasionally detected. It is proposed on the basis of these data that migration of antigen-stimulated IgG lymphoblasts, and perhaps of antigen, to spleen and peripheral lymph nodes may be dominant events after intestinal immunization of ruminants. This is consistent with the predominance of serum-derived IgG antibodies in colostrum and milk. Intramammary infusion of antigen gave rise to increases in antibody titers in all classes which were greater not only in lacteal secretions but also in blood serum than with either systemic route used. There was clear evidence from relative antibody titers for local synthesis of antibodies, principally IgA and IgG1, in the immunized glands. Comparison of IgA titers in secretions from the immunized glands with those in serum also suggested that locally synthesized IgA antibodies may have contributed in some measure to serum titers. Local synthesis in both immunized and nonimmunized glands was also reflected by the presence of increased numbers of IgA and IgG1 plaque-forming cells. It is hypothesized that antibody-forming cells responsible for local synthesis originated in lymphoid tissue within the mammary gland or from peripheral lymph nodes, depending upon the route of immunization.

Intestinal Goblet Cell Mucus Release

Abstract We previously reported that the infusion of certain soluble immune complexes stimulated mucus release from the rat small intestine in vivo. The present studies sought to evaluate the response of the intestine of normal and immunized rats to the infusion of antigen alone. One hour after the intraduodenal infusion of antigen, small intestinal washings were obtained and analyzed for the presence of 35S-labeled, high m.w. glycoprotein of goblet cell origin. The amount of goblet cell glycoprotein released was estimated from the radioactivity present in the void volume of a Sepharose 4B gel filtration column. The release of goblet cell mucus was enhanced by antigen stimulation in orally immunized animals. The discharge of goblet cell mucus was not increased after antigen infusion in animals immunized by the i.p. route despite the induction of high levels of serum antibody. The inability to demonstrate release of mucus after antigen challenge in systemically immunized rats suggests that the amount or the type(s) of antibody required at the mucosal surface is produced only after oral immunization.

Lung volumes after antigen infusion in the guinea pig in vivo: effects of vagal section.

The effects of intravenous antigen infusion on lung volumes and quasi-static deflationary pulmonary compliance in guinea pigs previously sensitized to ovalbumin were studied in vivo. Ovalbumin infusion significantly increased minimal gas volume to a similar extent in animals with intact or cut vagi. Total lung capacity fell only in animals with intact vagi. Quasi-static compliance fell in both groups of animals, but the fall was significantly greater in animals with intact vagi. These data demonstrate that immediate hypersensitivity reactions alter lung volumes and the elastic properties of the lung by both vagal dependent and vagal independent mechanisms.

Pulmonary response to antigen infusion in the sensitized guinea pig: modification by atropine.

Alterations in pulmonary conductance, dynamic compliance, respiratory frequency, minute volume, mean arterial pressure, pulse rate, relaxation volume-to-dry weight ratio, and wet-to-dry weight ratio resulting from antigen infusion in sensitized guinea pigs was examined with and without atropine treatment. In untreated animals 3 min after antigen infusion there were significant decreases in dynamic compliance and pulmonary conductance with an increase in relaxation volume-to-dry weight ratio while other parameters were not altered. In atropine-treated animals antigen infusion resulted in a decreased dynamic compliance and an increased relaxation volume-to-dry weight ratio but no significant change in pulmonary conductance. This suggests that the alterations in large and central airway tone resulting from antigen infusion are mediated predominantly by secondary cholinergic mechanisms while peripheral airway effects are mainly noncholinergic.

Transplantion resistance and potent antibody production induced in C3H/He mice against C3H/He mammary tumor by continuous infusion of solubilized antigen.

Transplantion resistance and potent antibody production induced in C3H/He mice against C3H/He mammary tumor by continuous infusion of solubilized antigen.

Local antigenic stimulation of guinea-pig mammary gland.

SummaryLocal immunization of the mammary glands of pregnant guinea‐pigs with killed Brucella abortus elicited a local production of specific antibody most of which was associated with IgA. The local response to intra‐ductal infusion of antigen was greater than that which followed injection of antigen into subcutaneous sites near the mammary gland. Local production of antibody persisted for two consecutive lactation periods following immunization by either route.Concentrations of IgA in milk were 10‐25 times greater than in serum, whereas the levels of IgG in milk were approximately 1/5th those for scrum. Significantly greater amounts of IgA were present in milk from locally‐infused' glands than in milk from glands immunized by subcutaneous injection and from non‐immunized glands. The levels of IgG in milk were not affected by local immunization.

Acute systemic anaphylaxis in the calf.

1. Acute systemic anaphylaxis in calves was characterized by marked systemic hypotension; hypertension in the pulmonary arteries and abdominal vena cava, and transient apnoea. Calves responded with a second reaction to antigen, but a third anaphylactic response could not be evoked.2. Suppression of systemic anaphylaxis could not be effected with mepyramine, whereas methysergide or diethylcarbamazine each suppressed anaphylaxis by 50%. Disodium cromoglycate alone did not inhibit anaphylaxis: however, when disodium cromoglycate was combined with diethylcarbamazine almost total suppression (85%) was achieved. Sodium meclofenamate also was a powerful inhibitor of anaphylaxis (80%). It is tentatively suggested that slow reacting substance (SRS-A) may be an important mediator of bovine anaphylaxis.3. Bilateral vagotomy did not modify the circulatory responses to injected histamine, 5-hydroxytryptamine (5-HT) or antigen, whereas the effects of these agents on ventilation (apnoea) were abolished by vagotomy.4. Plasma histamine concentration was increased during anaphylaxis, whereas plasma 5-HT was not. Whole blood histamine concentration fell sharply and remained depressed during 20 min of anaphylactic shock. Reduced whole blood histamine levels probably reflect the severe leucopoenia in the calves.5. Histamine concentrations in six tissues taken from calves subjected to anaphylaxis were not different from those in control calves; mast cells were of similar numbers to controls, but showed some swelling, granular spilling and metachromasia.6. Histamine, 5-HT, bradykinin and antigen caused increased pulmonary artery perfusion pressure and ventilation resistance in isolated lungs from sensitized calves. However, there was no difference in histamine and 5-HT concentration in perfusates obtained during antigen infusion of sensitized and control lungs.7. Systemic anaphylaxis of calves may result from the interaction of histamine, 5-HT and SRS-A. The present data implicate (by indirect measurement) SRS-A as an important mediator of anaphylaxis in this species.

Glomerular fibrin thrombi induced by antigen-antibody reactions: Protection by extreme thrombocytopenia

The role of platelets in the triggering mechanism of disseminated intravascular clotting induced by immune reactions was studied by comparing the effects of antigen in immunized rabbits and in immunized rabbits made extremely thrombocytopenic by the alkylating agent busulphan. The animals received an intravenous injection of Thorotrast 4 hr before the challenge with the antigen. The average platelet count prior to antigen infusion was 5000/μl in a group of 12 thrombocytopenic animals and 334,000/μl in a control group of 13 animals. Pretreatment with busulphan protected against glomerular fibrin thrombi and death provoked by the antigen. In comparison, 7 rabbits died in the control series, and fibrin thrombi were identified in the glomerular capillaries of 5 animals. We conclude that busulphan probably protects by depleting the animals of platelets, indicating that platelets are essential for the evolution of fibrin thrombi induced by immune reactions.

Properties of the histamine stores affected in the anaphylactic and anaphylatoxin shock of the guinea pig. I. Effect of pretreatment with anaphylatoxin.

1. Sensitized guinea pigs were protected with antihistamine and slowly injected (within 3 min) with antigen. 50% of the animals survived. After rapid injection of the same antigen dose (≙ 20 mg/kg ovalbumin) all animals died. 2. The lung histamine content was decreased to about half of the normal value in animals which died during the treatment as well as in surviving guinea pigs. 1 week later the lung histamine content was again within the normal range. 3. During this period an additional anaphylactic shock resulted only in a small reduction of the overall lethality (about 20%). Although no antihistamine had been given the animals died significantly later. Plasma histamine levels were depressed, sometimes to about 1/10 of the values found normally in anaphylactic shock. The lung histamine content did increase from 2 to 5 μg per g tissue. All parameters measured were normal 1 week after the antigen infusion. 4. During the same period of time the lethality of an anaphylatoxin shock was not reduced. Plasma histamine levels were diminished whereas the histamine content of the lungs did increase. As compared to an anaphylatoxin shock in unpretreated animals not exposed to antigen plasma histamine levels were in the normal range after 1 day. At this moment lung histamine content was still reduced by the antigen pretreatment. 5. We assume that the guinea pig lung contains two different histamine pools, one having a short half-life (less 1 day) and being depleted in anaphylatoxin shock, the other recovering more slowly (within 1 week), and being released in addition in anaphylaxis. The first one corresponds well with the properties of the non-mastcell-store described by Brodie et al. (1966), the latter seems to be identical with the typical mastcell-pool.

Properties of the histamine stores affected in the anaphylactic and anaphylatoxin shock of the guinea pig. II. Effect of pretreatment with antigen.

1. Sensitized guinea pigs were protected with antihistamine and slowly injected (within 3 min) with antigen. 50% of the animals survived. After rapid injection of the same antigen dose (≙ 20 mg/kg ovalbumin) all animals died. 2. The lung histamine content was decreased to about half of the normal value in animals which died during the treatment as well as in surviving guinea pigs. 1 week later the lung histamine content was again within the normal range. 3. During this period an additional anaphylactic shock resulted only in a small reduction of the overall lethality (about 20%). Although no antihistamine had been given the animals died significantly later. Plasma histamine levels were depressed, sometimes to about 1/10 of the values found normally in anaphylactic shock. The lung histamine content did increase from 2 to 5 μg per g tissue. All parameters measured were normal 1 week after the antigen infusion. 4. During the same period of time the lethality of an anaphylatoxin shock was not reduced. Plasma histamine levels were diminished whereas the histamine content of the lungs did increase. As compared to an anaphylatoxin shock in unpretreated animals not exposed to antigen plasma histamine levels were in the normal range after 1 day. At this moment lung histamine content was still reduced by the antigen pretreatment. 5. We assume that the guinea pig lung contains two different histamine pools, one having a short half-life (less 1 day) and being depleted in anaphylatoxin shock, the other recovering more slowly (within 1 week), and being released in addition in anaphylaxis. The first one corresponds well with the properties of the non-mastcell-store described by Brodie et al. (1966), the latter seems to be identical with the typical mastcell-pool.

Immune Response in the Mammary Gland

1.Three classes of immunoglobulins have been identified in lacteal secretions. IgG1 appears to be selectively transferred through the glandular epithelium into the udder. Presumably any method of immunization that stimulates circulating antibodies of the IgG1 class will be beneficial, resulting in IgG1 antibodies in the milk.2.IgA class immunoglobulins are also found in the milk, however, in lower concentration than IgG1 immunoglobulins. IgA is produced locally in the gland since an increase is seen in antigen-infused quarters over noninfused quarters. A factor that might be important is that probably the total amount of any immunoglobulin in lacteal secretions will not be increased by systemic immunization whereas local immunization does increase the amount of IgA.3.The method and route of immunization is possibly very important. Intramuscular, subcutaneous, or other routes that result in systemic stimulation of antibodies, to be effective, must for the most part stimulate class IgG1 immunoglobulins. Hammer, Kickhofen, and Henning (7) have recently published that there is a quantitative shift of antibody activity from IgG1 to IgG2 between the first and third courses of immunization of cattle (7). These IgG2 antibodies would be of little value for mammary gland immunity insofar as antibodies in the milk are concerned. However, since the tissues of the mammary gland are involved in a mastitis infection, specific antibodies of all immunoglobulin classes may be beneficial. Immunization by inoculation of a Streptococcus agalactiae, type II preparation into the area of the supramammary lymph node stimulates protective antibodies in the milk. These antibodies have been shown to be of IgG class principally, with less IgA. Finally, by intramammary infusion of antigen into one or more quarters of a nonlactating gland, an increase in specific antibodies of only the IgA class was noted.4.Progress in the determination of the immunogenic substance necessary to stimulate resistance to staphylococcal mastitis has been slow. Serological typing of staphylococci is not available; however, on the basis of phage typing, most mastitis staphylococci may fall into a dozen or so types (14). Serological typing and the immunogenic response of the group B streptococci are much further advanced. The immunology of the group C and group E streptococci is also in a primitive state. It probably is not necessary to separate and purify each immunogen that is required in a preparation; however, it is necessary that inactivated bacteria of each different antigenic type be present in a bacterin in order to stimulate all the various antibodies necessary.One can speculate that a combination of immunization routes that result in the stimulation of several of the classes of immunoglobulins might produce sufficient protective antibodies in the milk and in the mammary gland tissues to increase resistance. Local stimulation would increase specific IgA while systemic stimulation would result in specific IgG1, IgG2 and IgM. Local stimulation would presumably have to occur in the nonlactating gland with a nonirritating or nontissue damaging antigen. The antigen may be a bacterin consisting of a combination of each of the types of staphylococci and streptococci and be inactivated with formalin or some other inactivant. The bacterin should also contain staphylococcal toxoids.

Antibody-producing cells in antigenically stimulated mammary glands and in the gastro-intestinal tract of sheep.

SummaryThe histological changes in normal mammary glands of ewes during the period one month before and after lambing have been compared with the changes occurring in glands infused with antigen one month before lambing. Attention was focused on the changes in concentration and on the location of cells of the lymphocyte‐plasma cell and macrophage‐monocyte series.The most significant feature following antigen infusion was the striking appearance of pyroninophilic cells usually situated in areas adjacent to the glandular epithelium. Lymphocytes were also more numerous in infused than non‐infused glands until early lactation.Attempts were made to detect specific immunoglobulin‐producing cells in mainmary glands, certain other glandular tissues and in lymph nodes. When sections of infused mammary glands were stained with specific antisera conjugated with fluoresce in isothiocyanate numerous fluorescing cells were observed and their location correlated closely with that of the pyroninophilic cells seen in paraffin sections. The majority of the fluorescing cells were of IgA‐specificity with IgG1‐ and IgM‐specific cells present in lower concentrations. In the gastro‐intestinal tract fluorescing cells located in the lamina propria were abundant and the majority were IgA‐specific. Numerous IgG1‐ and IgG2‐specific cells were present in the lymph nodes. Very few pyroninophilic or flourescing cells were seen in sections of the parotid salivary gland, uterus and vagina.The observations indicate that the majority of pyroninophilic cells in antigen stimulated mammary glands and in the gastro‐intestinal tract are IgA‐specific cells which are responsible for the local production of antibody.

Carotid baroreceptor activity during antigen infusion in the sensitized cat.

ARTICLESCarotid baroreceptor activity during antigen infusion in the sensitized catRS Tuttle, and M McClearyRS Tuttle, and M McClearyPublished Online:01 Jun 1969https://doi.org/10.1152/ajplegacy.1969.216.6.1314MoreSectionsPDF (1 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation More from this issue > Volume 216Issue 6June 1969Pages 1314-1321 Copyright & PermissionsCopyright © 1969 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1969.216.6.1314PubMed5786717History Published online 1 June 1969 Published in print 1 June 1969 Metrics

The Immune Response of the Bovine Species Infused Intramammarily with a Semi-Purified Preparation of the Flagella of Salmonella Bacteria

Summary Using the methods of immobilization and bacterial chaining (somatic agglutination) for the determination of Salmonella anti-H and anti-O titer respectively, the cow was found to have an initial blood serum anti-H titer of 10 to 80 and anti-O titer of 20 to 320. Similar initial anti-H and anti-O titers were not always found in the milk. Infusion of the mammary gland with a semi-purified preparation of Salmonella flagella brought about an acute inflammation of the gland, a high systemic temperature, a drop in milk production and a shift to a more alkaline pH of the milk. An anti-H and anti-O titer response was stimulated in both the blood serum and the milk. The titer of the blood serum was invariably higher than that of the milk and the anti-O response was dominant to the anti-H response. The greatest increase in titer followed within 6 to 10 days of the first antigen infusion. Subsequent antigen infusions only maintained the response from the first infusion. The titer of the blood remained much more constant than that of the milk with the milk returning to initial titer levels within 7 to 8 days postinfusion. There was a definite titer response in the milk of the noninfused quarters; however, it was not as marked or as persistent as the titer of the infused quarters. Intramammary infusion of a semi-purified preparation of Salmonella flagella brought about a definite systemic antibody response. The marked anti-O titer indicates that part of the cellular somatic antigen remains attached to the flagella when they were removed from the cell wall. In the blood serum the anti-O antibody was predominantly IgG while anti-H antibody was about 50% IgG and 50% IgM. Semi-purified flagellar antigen of Salmonella infused intramammarily at a level of 1.0 µg/kg body weight elicited antibody response, but a dose of 9.71 µg/kg suppressed antibody response. With intravenous injection of the same antigen preparation in rabbits, a higher anti-H titer was obtained than anti-O titer.

Local production of antibody by the lactating mammary gland following antigenic stimulation.

SummaryExperiments have been carried out to determine whether the infusion of antigens into the dry mammary glands of pregnant ewes gives rise to a sustained local immunity. One side of the udder was infused with killed Salmonella typhi “O” organisms and the other with a similar dose of killed Brucella abortus organisms. Infusions were given to 6 pregnant ewes about 1 month before parturition.Although the anti‐Salmonella and anti‐Brucella titres in the colostrum from both sides were higher than those in plasma, the titres of specific antibody in colostrum from the sides receiving corresponding specific antigen were usually higher than in the colostrum from the opposite side. These results indicated that both local synthesis and active transfer of antibody were occurring at this time.There was a rapid fall in the antibody titres of milk within the first week of lactation. However, the ratio of the titres of specific antibody in the samples from the sides receiving corresponding specific antigen to those in plasma were usually well above unity throughout lactation. On the other hand, the milk: plasma ratios computed from the anti‐Salmonella and anti‐Brucella titres in plasma and in whey samples collected from the sides which were not infused with corresponding antigens rapidly decreased within 2 weeks after parturition to values which varied between 0·03–0·25. These results indicated that during lactation most of the specific antibody in milk was of local origin.The anti‐Salmonella and anti‐Brucella titres in the mammary lymph from the sides infused with Salmonella and Brucella respectively were lower than those in the milk from the same side. In view of the relative impermeability of the glandular epithelium of the mammary gland to γ‐globulin molecules, this finding suggests that the cells responsible for local synthesis of antibody are in close proximity to the glandular epithelium.