Lymphocyte blastogenic responses to inciting food allergens in dogs with food hypersensitivity

Lymphocyte blastogenic responses against food allergens in dogs with food hypersensitivity were evaluated in this study. Eleven dogs with food hypersensitivity, based on food elimination and oral food provocation tests and allergic responses to food allergens, were examined by various tests such as intradermal testing, antigen-specific IgE testing, and lymphocyte blastogenic responses. The number and kinds of food allergens identified as positive by these tests were compared with the offending food allergens that were found in an oral food provocation test. In 9 (82%) of the 11 dogs with food hypersensitivity, there was close agreement for positive allergens between the results of lymphocyte blastogenic responses and oral food provocation test; however, there was little agreement for intradermal and IgE testing of the positive allergens with those of the oral food provocation test (11% and 31%, respectively). In the 9 dogs, the stimulation indices of lymphocyte blastogenic responses increased to 2.0-10.1 upon food provocation but decreased significantly to 0.7-1.4 upon feeding the elimination diet until clinical signs disappeared. These results indicate that lymphocyte blastogenic responses may fluctuate because of exposure to offending food allergens in dogs with food hypersensitivity. Lymphocytes reactive to food allergens may play an important role in the pathogenesis of food hypersensitivity in dogs.     

Experimentally induced Cooperia oncophora infection in calves: lymphocyte blastogenic and delayed hypersensitivity responses

Lymphocytic responses in peripheral blood and visceral lymph to Cooperia oncophora antigen and skin tests were determined in 35 Holstein male calves that were inoculated orally with single or multiple doses of C oncophora infective larvae. Several calves were vaccinated or given immune serum before larvae were inoculated. Antigen-specific in vitro blastogenesis of blood and lymph lymphocytes and delayed-type hypersensitivity reactions were observed in several inoculated, vaccinated, and/or passively immunized calves. Most calves that had delayed skin reactions also had in vitro lymphocyte responses to C oncophora antigen. The lymphocyte and skin responses were inconsistent and variable in time of onset--the earliest lymphocyte response occurring 7 days after calves were inoculated. A cellular immune response was induced by both dermal vaccination and oral inoculation; however, passive immunization by IV administration of immune serum simultaneously with inoculation did not have an apparent effect on the cellular response, as measured by the lymphocyte blastogenesis test or dermal testing. Although cellular immune responses were observed in several calves infected with C oncophora, there was no apparent relationship between the specific responses and number of nematodes establishing infection in calves after either single- or multiple-dose oral inoculations.     

Changes in cell-mediated immune responses after experimentally-induced anaphylaxis in dogs.

Experimentally-induced type 1 hypersensitivities were induced in normal dogs to either ovalbumin or Ascaris antigen. In vitro and in vivo cell-mediated immune responses were measured before sensitization and again at 1 and 6 days after induction of anaphylaxis by intravenous challenge with antigen. Histamine-modulated lymphocyte functions, such as histamine-induced suppression, histamine co-mitogen induced blastogenesis and the in vivo cutaneous responses to intradermally injected mitogens decreased post anaphylaxis. Spontaneous suppression of the autologous mixed-lymphocyte reaction increased post anaphylaxis. Lymphocyte blastogenic response to Concanavalin A (Con A) decreased at 6 (but not at 1) days post anaphylaxis probably due to a mediator other than histamine. Blastogenesis of 24 h preincubated cells by suboptimal concentration of Con A, declined post anaphylaxis, but Con A-induced suppression was not significantly altered. Dogs with atopic dermatitis have some altered cell-mediated immune responses. Altered histamine-induced and spontaneous suppression, histamine suppression of mitogenesis and decreased contact sensitivity observed in this experimental type 1 hypersensitivity mimicked that of atopic dogs. Increased cutaneous response to mitogens observed in atopic dogs was not reproduced in the type 1 hypersensitive dogs. These findings suggest some of the altered cell-mediated immune functions observed in dogs with atopic dermatitis result from type 1 hypersensitivity. The other abnormalities may be intrinsic to the atopic state.     

Lymphocyte blastogenic responses to food antigens in cats showing clinical symptoms of food hypersensitivity

Three cats were diagnosed as having food hypersensitivity by food elimination and oral food provocation tests. Twelve allergenic food ingredients were identified by oral food provocation test in the 3 cats. Of the 12 food ingredients, 9 offending food antigens were shown to be positive in a lymphocyte stimulation test; however, none of them were positive in antigen-specific IgE testing, and only four food antigens were positive in intradermal testing. The stimulation indices in the lymphocyte stimulation tests for the 9 food ingredients were found to be decreased after the cats were fed elimination diets. The present study demonstrates that the lymphocyte stimulation test reflects an immunologic reaction involved in food hypersensitivity and can help identify allergenic food ingredients in feline food hypersensitivity.     

Relationship between mitogen receptors in peripheral blood lymphocytes and blastogenic responses to mitogen

The relationship between the optimum concentration of mitogen which induces lymphocyte blastogenic response and the receptor occupancy by mitogen was investigated. The receptor occupancies which induced maximal blastogenic activity in equine, bovine and canine peripheral blood lymphocytes (PBL) were 31.1 per cent, 26.5 per cent and 38.4 per cent with phytohaemagglutinin-P, and 48.2 per cent, 17.9 per cent and 24.5 per cent with concanavalin A, respectively. The data clearly show that each animal species had its own optimum concentration of mitogen for stimulation of PBL. Optimum concentration for blastogenesis and number of binding sites of each mitogen had a good correlation with each other for all three species.     

Comparison of the blastogenic response of peripheral blood lymphocytes from canine parvovirus-positive and -negative outbred dogs

Lymphocyte blast transformation assays (LBT) were performed on canine parvovirus (CPV) -positive and -negative mongrel dogs randomly selected from a humane facility. Concanavalin A as well as Phytohemagglutinin P stimulation was depressed (p less than 0.001) in the group of animals shedding CPV compared to CPV-negative dogs.     

DNA vaccination against Japanese cedar pollinosis in dogs suppresses type I hypersensitivity by controlling lesional mast cells

Sensitization to allergens of Japanese cedar pollen is known to cause canine atopic dermatitis as approximately 10% of atopic dogs in Japan were positive to the pollen allergen. Among the two major allergens of Japanese cedar pollen, since Cry j 1 is more important than Cry j 2 as an antigen to increase IgE in atopic dogs sensitized to Japanese cedar pollen, Cry j 1 can be a target for immunotherapy. In our study, efficacy of DNA vaccination with a plasmid containing the gene of a major allergen of Japanese cedar (Cryptomeria japnonica, CJ) pollen, Cry j 1, was examined using a dog model experimentally sensitized to CJ pollen allergen. Cry j 1 DNA plasmid and a vector plasmid (pCAGGS) were injected into six dogs and three dogs, respectively, five times with an interval of 1.5 month. After the treatment with Cry j 1 DNA plasmid, production of IgE against Cry j 1 decreased in four of the six dogs in the treatment group, whereas it increased in the three dogs of the control group. The reactivity to the pollen allergen in intradermal testing and provocation testing were obviously reduced in the treatment group, but not in the control group. The number of mast cells in alveolar area of the lung in the treatment group was smaller than that in the control group. Cry j 1 DNA plasmid was also injected into three atopic dogs sensitive to Cry j 1, resulting in improvement of clinical signs in the pollination season. These findings indicated that Cry j 1 DNA plasmid could regulate mast cell-mediated reaction against Cry j 1, which could be an alternative and effective treatment for CJ pollinosis.     

Effects of the lipopolysaccharide-protein complex and crude capsular antigens of Pasteurella multocida serotype A on antibody responses and delayed type hypersensitivity responses in the chicken.

The effects of the lipopolysaccharide-protein complex (LPS) and crude capsular antigen (CCA) prepared from Pasteurella multocida serotype A isolated from a duck in the Philippines, on antibody responses to sheep red blood cells (SRBC) and Brucella abortus (BA) and delayed type hypersensitivity (DTH) responses to bovine serum albumin (BSA) in the chickens were studied. Chickens injected subcutaneously with LPS and CCA at 1 and 2 weeks of age and immunized intravenously with the mixed antigens of SRBC and BA, at 3 and 4 weeks of age showed significantly increased antibody responses against both SRBC and BA, when evaluated at 7 days after each immunization. In addition, these chickens sensitized intramuscularly with the emulsion of BSA in complete Freund's adjuvant at 5 weeks of age, and then injected into the wattle with BSA at 7 weeks of age also showed significantly increased DTH responses against BSA, when evaluated at 24 and 48 hr after challenge. These results indicate that LPS and CCA of P. multocida serotype A have a property enhancing humoral and cell-mediated immune responses.     

In vitro blastogenic responses of peripheral lymphocytes from marmosets to phytohemagglutinin and to autologous lymphocytes transformed by Epstein-Barr virus

White-lipped marmosets were evaluated for their cell mediated immune (CMI) response to EBV to determine the feasibility of CMI studies in marmoset models for EBV oncogenesis. The mitogen, cell concentrations, the length of incubation period and serum requirements were defined for in vitro lymphocyte stimulation tests. The level of response of each animal was dependent on the concentration of phytohemagglutinin-P (PHA-P) and was independent of cell densities employed. The rate of tritiated-thymidine incorporation by mononuclear cells due to PHA-P increased exponentially between 2–4 days. This test was reproducible for a given batch of PHA-P when the cells were cultured in the presence of 10% heat inactivated fetal bovine serum. The five white-lipped marmosets were seronegative for EBV antigens and did not show lymphocyte stimulation with EBV particles and EBV soluble antigen, but two of these animals exhibited significant stimulation with autologous lymphocytes transformed in vitro by B95-8 virus. Despite the limited amount of blood (3–4 ml) that could be obtained from each animal in a single bleeding, it was possible to perform multiple lymphocyte stimulation assays with the protocol used.     

Immediate type hypersensitivity in dogs induced by storage mites

Results of intradermal skin tests on 53 dogs, employing storage mite extracts as antigen, indicated that storage mite allergy is fairly common. Only four of 29 dogs exhibiting no allergic symptoms showed skin reactions to the most common storage mite, Acarus siro, whereas 18 out of 24 dogs with symptoms assumed to be associated with allergy showed such reactions. The passive cutaneous transfer test revealed that an immediate type hypersensitivity reaction was involved.     

Platelet aggregation in dogs after live-virus vaccination

Platelet aggregation induced by ADP was studied in dogs after vaccination with a modified live-virus distemper vaccine. A significant decrease in platelet counts was observed about 1 week after the vaccination. There were no consistent changes in the aggregability of the platelets. In 4 samples the aggregability was significantly increased compared to the prevaccination values. The observed changes will not cause a defective hemostasis in otherwise normal dogs.     

Duration of immunity in dogs after vaccination or naturally acquired infection.

This paper reviews current scientific information about the duration of immunity induced in dogs by infection or vaccination. It describes the shortcomings of the methods used to measure the immune responses of dogs, and explains the need for basic studies on the nature of protective humoral and cellular responses, and standardised assays for the long-term duration of immunity to pathogens other than rabies. The information is inadequate to warrant uniform recommendations on the ideal intervals for vaccination; each vaccine must be evaluated on the basis of its own merits and the characteristics of the disease it is intended to guard against.     

Anaphylaxis after rabies vaccination for dogs in Japan

Severe adverse reactions after rabies vaccination in dogs were examined from 317 cases reported to the Ministry of Agriculture, Forestry and Fisheries (MAFF) in Japan during 15-year period from April 2004 to March 2019. We found that 109 of the 317 dogs showed anaphylaxis (0.15/100,000 vaccinated dogs), and 71 of the 109 cases of anaphylaxis resulted in death (0.10/100,000 vaccinated dogs). We measured bovine serum albumin (BSA) in four commercially available rabies vaccines and found the levels ranged from 0.1 to 16.6 µg/dose. Our survey showed that the rate of anaphylaxis to rabies vaccines in dogs is rare, although some cases of anaphylaxis resulted in death. Veterinarians should be well prepared to deal with vaccine-associated anaphylaxis.     

Influence of sarcoptic mange and cold and ambient temperature on blastogenic responses of lymphocytes and serum cortisol concentrations of pigs

Blood samples from sarcoptic mite-infested pigs were evaluated for effects of mite infestation and cold and ambient temperatures on lymphocyte blastogenic responses and for effects of mite infestation on serum cortisol concentrations. In experiment 1, sarcoptic mite-infested and noninfested pigs were housed in cold (5 to 15 C fluctuating) and thermoneural (25 C) environmental chambers for 5 weeks. Differences were not observed (P greater than 0.10) in blastogenic responses to phytohemagglutin or pokeweed mitogen between lymphocytes from infested and noninfested pigs on postinfestation days (PID) 7, 21, 28, and 35 in either environmental chamber. When lymphocytes from noninfested pigs were cultured with sera from infested pigs, alterations of blastogenic responses were not detected. Cortisol values were higher (P less than 0.05) in sera from sarcoptic mite-infested pigs, compared with those from noninfested pigs, at 4 PM on PID 14 and 4 AM and 10 AM on PID 15. Cortisol values were higher (P less than 0.05) in sera obtained at 10 AM on PID 14 and at 10 AM on PID 15 from pigs housed in cold chambers, compared with those from pigs housed in thermoneutral chambers. Interactive effects between sarcoptic mite infestation and cold ambient temperatures were not observed. At 4 AM on PID 15 (experiment 2), cortisol values were higher (P less than 0.05) in sera of infested pigs, compared with those in noninfested pigs. Seemingly, sarcoptic mange in pigs did not alter mitogen-induced lymphocyte blastogenic responses, but did increase serum cortisol concentrations, indicating that sarcoptic mange may be a stressor in pigs.     

Effect of sodium diethyldithiocarbamate, Corynebacterium parvum and mycobacterium cell wall extract on in vitro blastogenic responses of bovine blood lymphocytes

In vivo inoculation of three-month-old calves with sodium diethyldithiocarbamate (DTC), killed Corynebacterium parvum or mycobacterium cell wall extract (MCWE) resulted in an enhancement of in vitro peripheral blood lymphocyte blastogenic responses to mitogens phytohemagglutinin (PHA) and Concanavalin A (Con A) in the first three days after treatment. In a separate experiment, blood lymphocytes isolated from a healthy nontreated calf were incubated in vitro in presence of each of the same immunostimulating agents and tested for their blastogenic responses to PHA and Con A. The results showed that all immunostimulants, excepting DTC, enhanced the in vitro blastogenic responses of lymphocytes to PHA and Con A. Finally, addition of MCWE to cultures of blood lymphocytes isolated from calves vaccinated intramuscularly with bovine rotavirus and adjuvant resulted in an enhancement of the in vitro lymphocyte transformation to rotavirus. Our study demonstrated that DTC, killed Corynebacterium parvum and mycobacterium cell wall extract were able to enhance bovine T cell proliferation in vitro.     

Dietary hypersensitivity in cats and dogs

Adverse reactions to food or dietary hypersensitivity are frequently seen problems in companion animal medicine and may be difficult to differentiate from inflammatory bowel disease (IBD). Dietary hypersensitivity can be divided into two subgroups: immunological and nonimmunological problems. Non-immunological problems can be subdivided into food intolerance, food poisoning, and dietary indiscretion. The immunological group can be subdivided into true food allergy (IgE mediated) and anaphylaxis (non-IgE mediated). This article gives an outline of what dietary hypersensitivity is, and more specifically food allergy and how to deal with patients with possible dietary hypersensitivity.     

Immunologic responses against hydrolyzed soy protein in dogs with experimentally induced soy hypersensitivity

OBJECTIVE: To assess whether dogs with experimentally induced type I hypersensitivity against soy protein would respond to soy hydrolysate and develop cutaneous or gastrointestinal tract reactions after intradermal and oral challenge exposure. ANIMALS: 12 na?ve Beagle pups (9 sensitized and 3 control dogs). PROCEDURE: 9 dogs were sensitized against soy protein by administration of allergens during a 90-day period. After the sensitization period, serum concentrations of soy-specific IgE were determined and an intradermal test was performed to confirm the dogs were sensitized against soy protein. An intradermal challenge test and an oral challenge test with native and hydrolyzed soy protein were conducted on 6 sensitized and 2 control dogs. RESULTS: High serum concentrations of soy-specific IgE and positive results for the intradermal test were observed for the 9 sensitized dogs after completion of the sesitization process. Sensitized dogs challenge exposed with hydrolyzed soy protein had a reduced inflammatory response after intradermal injection and no clinical response after an oral challenge exposure, compared with responses after intradermal and oral challenge exposure with native soy protein. CONCLUSIONS AND CLINICAL RELEVANCE: Soy-sensitized dogs did not respond to oral administration of hydrolyzed soy protein. Thus, hydrolyzed soy protein may be useful in diets formulated for the management of dogs with adverse reactions to food.