Evaluation of systemic and secretory IgA concentrations and immunohistochemical stains for IgA-containing B cells in mucosal tissues of an Irish setter with selective IgA deficiency

Immunoglobulin A is the predominant secretory antibody at mucosal surfaces. In the dog, immunoglobulin A deficiency (IgAD) is characterized by low to absent serum IgA and normal to elevated serum immunoglobulin G (IgG) and immunoglobulin M (IgM) concentrations. However, studies comparing serum and secretory IgA in dogs have often documented a poor correlation, suggesting that serum concentrations should not be used to estimate mucosal secretion of this antibody. This report demonstrates the concurrent use of serum IgA, IgG, and IgM; secretory IgA (from bronchoalveolar lavage fluid); and immunohistochemical stains on bronchial and duodenal mucosa for IgA-containing B cells in a young Irish setter with recurrent respiratory and gastrointestinal signs.     

Serum concentrations of IgG, IgA, and IgM in retired racing Greyhound dogs

Background: Greyhound dogs have significant physiologic, hematologic, and biochemical differences when compared with other breeds, including significantly lower serum globulin concentration owing to decreases in the α‐ and β‐globulin fractions. The specific proteins that account for differences in globulin concentrations are not known, but IgA and IgM, both β‐globulins, are potential candidates. Objectives: The aims of this study were to measure serum IgG, IgA, and IgM in clinically healthy retired racing Greyhounds and compare the results with those of age‐ and sex‐matched non‐Greyhound dogs. Methods: Study animals included 25 Greyhound and 20 non‐Greyhound dogs. Total protein, albumin, and total globulin concentrations were determined. IgG, IgA, and IgM concentrations were measured using a commercially available radial immunodiffusion kit. The Student t‐test assuming equal variances was used to compare concentrations of immunoglobulins between groups. Results: Serum concentrations of IgA and IgM in Greyhounds (IgA=49±20 mg/dL; IgM=132±47 mg/dL) were significantly lower than concentrations in non‐Greyound dogs (IgA=70±39 mg/dL; Ig M=212±78 mg/dL). Concentrations of IgG did not differ between groups. Conclusions: Mean serum IgA and IgM concentrations in Greyhounds were lower than those in non‐Greyhound dogs. This may contribute to low serum concentrations of β‐globulins in Greyhounds. Specific reference intervals are recommended for Greyhounds to avoid possible misdiagnosis of IgA or IgM deficiency.     

Relative IgA deficiency and small intestinal bacterial overgrowth in German shepherd dogs

Serum immunoglobulin concentrations and densities of IgA-producing immunocytes in intestinal mucosa were compared in a group of clinically healthy dogs of various breeds, a group of clinically healthy German shepherd dogs, and a group of German shepherds with bacterial overgrowth in the proximal small intestine. Serum concentrations of IgA, but not IgM or IgG, were significantly lower in the clinically healthy German shepherd dogs than in other purebreed and mixbreed dogs, indicating that production of IgA by gut-associated lymphoid tissue might be relatively low in this breed. However, densities of IgA-producing cells were not significantly different comparing these two groups, suggesting that any impairment of mucosal IgA production is more likely to be related to defective synthesis or secretion of IgA than to reduced numbers of IgA-producing immunocytes. Comparable findings in German shepherd dogs with small intestinal bacterial overgrowth provided further indirect evidence that local immunity might be defective in this breed, since these luminal bacteria would be expected to stimulate mucosal IgA production. However, it is not clear whether such a defect is directly responsible for the overgrowth, or whether there is an indirect relationship between defective local immunity and bacterial overgrowth in German shepherd dogs.     

Quantitation of the immunoglobulins in reproductive tract secretions of the mare

IgG, IgA, IgM and albumin concentrations were measured in serum, follicular fluid and oviductal, uterine and intestinal secretions of the horse. Follicular protein concentrations were found to be dependent on serum concentration and molecular size. Of the immunoglobulins only IgG was detectable in oviductal secretions, but IgG:albumin ratios did not differ significantly from those in serum. IgG, IgA and IgM were measured in uterine secretions, with IgG predominant. Serum transudation into uterine secretions was minimal. In intestinal secretions, IgA levels were slightly higher than IgG, with albumin and IgM at low levels. In five mares with histories of chronic metritis, IgG, IgA and albumin concentrations were significantly elevated in uterine secretions.     

Serum concentrations of immunoglobulins G, A, and M in dogs with neoplastic disease

The concentrations of the three major classes of immunoglobulins (Ig) were determined in canine serum by single radial immunodiffusion against calibrated standards. Serum samples were collected from 121 dogs categorized into 3 groups: normal dogs (n = 34), those with lymphosarcoma (n =41), and those with malignant, solid neoplasms other than lymphosarcoma (n = 46). The mean value for serum IgM concentration in the group of dogs with lymphosarcoma was significantly (P less than or equal to 0.02) higher than was the mean IgM concentration of the normal dogs. Dogs with neoplasms other than lymphosarcoma had significantly increased serum concentrations of IgG and IgM antibodies. There was no significant difference in serum IgA concentration among the three groups. A wide range of Ig concentrations was in the serum of clinically normal dogs, as well as in dogs with neoplastic diseases. Although individual dogs with neoplasms had low serum Ig concentrations, the 81 dogs with neoplastic disease were generally able to synthesize Ig.     

Survey of serum IgA, IgG, and IgM concentrations in a large beagle population in which IgA deficiency had been identified

Concentrations of serum IgA, IgG, and IgM were determined for 829 adult Beagles from a commercial kennel in which several IgA-deficient dogs had been identified previously (index kennel). These values were compared with measurements of 100 adult dogs from another Beagle kennel (control kennel). After adjustment for differences in the ages and gender of the dogs, dogs from the index kennel had significantly (P less than 0.0001) lower IgA concentrations (mean, 46 mg/dl) than dogs from the control kennel (mean, 68 mg/dl). Regardless of kennel, males had significantly (P less than 0.01) higher IgA concentrations than did females. Dogs in the control kennel had significantly (P less than 0.04) higher IgG concentrations (mean, 2,649 mg/dl) than did dogs in the index kennel (mean, 2,478 mg/dl), and female dogs in the control kennel had significantly (P less than 0.05) higher IgM concentrations (mean, 189 mg/dl) than dogs of either sex in the index kennel (mean, 162 mg/dl) or male dogs in the control kennel (mean, 163 mg/dl). For both sexes, concentrations of IgA, IgG, and IgM increased with age.     

感染鸡贫血病毒雏鸡接种新城疫疫苗后局部粘膜免疫功能的变化

雏鸡1日龄感染鸡贫血病毒（CAV），8日龄接种Lasota疫苗，以未感染免疫雏鸡为对照，于免疫后7、14、28d检测其哈德尔腺和盲肠扁桃体T细胞及IgG^ 、IgM^ 、IgA^ 抗体生成细胞数量，泪液、气管液、肠液、胆汁中IgG、IgM、IgA含量以及泪液、胆汁HI抗体滴度的动态变化。揭示了感染CAV雏鸡接种ND疫苗免疫后哈德尔腺、盲肠扁桃体的T细胞和IgG^ 、IgM^ 、IgA^ 抗体生成细胞数量，泪液、气管液、肠液、胆汁中免疫球蛋白IgG、IgM、IgA含量以及泪液、胆汁HI抗体滴度，均较未感染免疫雏鸡明显减少。表明眼部、呼吸道和消化道局部粘膜免疫防御能力减弱。     

Systemic and local intestinal antibody response in dogs given both infective and inactivated canine parvovirus

Systemic and local immune responses were evaluated in dogs given infective canine parvovirus (CPV) and 2 administrations of inactivated CPV 6 months later. Before the inactivated CPV was given, a jejunal cannulation was performed on the animals. During infective CPV administration, concentrations of class-specific copro- and sero-immunoglobulin (Ig)G, IgA, and IgM were determined by enzyme-linked immunosorbent assay. High concentrations of both copro- and sero-IgM, as well as a moderate increase in the concentrations of sero-IgG and copro-IgA, were detected within 3 days after experimental challenge. Hemagglutination inhibition titers correlated with both serum anti-CPV IgG and IgM early and serum anti-CPV IgG in the later states of infection. After 2 oral administrations of inactivated CPV, class-specific jejuno-, copro-, and sero-IgG, IgM, and IgA anti-CPV antibodies also were determined by enzyme-linked immunosorbent assay. High concentrations of jejuno-IgM and moderate levels of jejuno-IgG and IgA were found. Copro-IgM was not detected in the feces; however copro-IgG and IgA were. Also, no correlations were found to exist when sero-IgM and IgG were compared with jejuno- and coproantibody concentrations throughout the experimental period. Thus, it appears that the immune responses to CPV include both a secretory component of the intestinal mucosa and a systemic component of peripheral lymphoid tissues. Application of the modified Witzel's enterostomy to this study of local intestinal immunity proved beneficial. The technique was well tolerated by experimental animals and allowed for simple, multiple sample collections of intestinal contents for virus and antibody determinations with apparently minimal alterations in the luminal environment.     

Quantitative Changes Associated with Calving in the Levels of Bovine Immunoglobulins in Selected Body Fluids I. Changes in the Levels of IgA, IgG1 and Total Protein

Levels of bovine IgA, IgG1 and total protein (TP) were determined in serum, saliva, tears and individual quarter lacteal secretions of six Holstein-Friesian cows sampled from six weeks before to four weeks after parturition. Hierarchal analyses of variance indicated significant variations among weeks, cows and quarters of the udder. A precipitous but non proportional drop in the levels of IgA and IgA1 in lacteal secretions occurred at calving. There was a concomitant increase in IgG1, and decrease in IgA, in serum. Correlation studies supported the concept of selective transport of IgG1 from serum to lacteal secretions in regulated amounts independent of serum IgG1 levels. Changes in the IgG1/TP ratio of serum and lacteal secretions supported the idea of a decrease in the selective transport mechanism. Correlation studies and estimations of secretory IgA (SIgA) in serum suggest that serum IgA is derived from IgA synthesized in secretory tissues. Highly significant correlations between IgA and IgG1 levels in all secretions postpartum suggest that local IgA synthesis and either IgG1 transport or local IgG1 synthesis are initiated by the same stimuli. Although some of the variation in the level reported for IgA and IgG1 in secretions resulted from protein dilution, much of the variation represents physiological differences between individual animals and tissues in the same animal. An IgG2/IgG1 ratio approaching that of serum occurred in a mastitic quarter of one cow. IgA was the principal immunoglobulin in saliva and tears, comprised a greater proportion of the immunoglobulin in milk whey than in prepartum lacteal secretions and was a minor immunoglobulin in bovine serum.     

Profile of anti-Leishmania antibodies related to clinical picture in canine visceral leishmaniasis

This research investigated the profile of anti-Leishmania antibodies in different clinical forms of canine visceral leishmaniasis (CVL). Naturally infected dogs were divided into two groups: subclinical dogs (SD, n=10) and clinical dogs (CD, n=68). Non-infected dogs (ND, n=7) comprised the negative control group. The humoral response was evaluated by the profile of total IgG, IgG1, IgG2, IgM, IgA and IgE, determined by ELISA. Infected animals showed increased levels of total IgG, IgA and IgE in addition to IgG1 and IgG2 in groups SD and CD, when compared with group ND. Furthermore, it was observed that IgG2 and IgM were correlated with symptomatology, while total IgG, IgG1 and IgA were negatively correlated and IgE showed no correlation. It follows that serum levels of IgG2 anti-Leishmania are correlated with typical clinical signs of disease. Furthermore the determination of specific anti-Leishmania antibodies could be an important tool in monitoring CVL clinical picture.     

Immunoglobulin concentrations in ovine body fluids.

Ovine IgG, IgM and IgA and antisera specific for these immunoglobulins were prepared. The specific antisera were used to estimate the immunoglobulin concentrations in certain sheep body fluids. IgA was shown to be the major immunoglobulin in saliva, lung and lachrymal fluid, tracheobronchial and nasal secretions while IgG was the predominant immunoglobulin in colostrum, milk, bile and serum.     

Salivary and serum immunoglobulin levels in cats with chronic gingivostomatitis

The salivary and serum concentrations of immunoglobulins G, M and A (IgG, IgM and IgA), and the salivary concentrations of albumin were measured by ELISA in 30 cats with chronic gingivostomatitis and 32 healthy cats. The cats with chronic gingivostomatitis had significantly higher salivary concentrations of IgG, IgM and albumin, and higher serum concentrations of IgG, IgM and IgA, but significantly lower salivary concentrations of IgA than the healthy cats. The cats with chronic gingivostomatitis were treated with either methylprednisolone, sodium aurothiomalate, metronidazole and spiramycin, or oral hygiene products. After three months of treatment, the cats receiving methylprednisolone had a significant reduction in serum IgG levels compared to the cats treated with sodium aurothiomalate or metronidazole and spiramycin, but after six months of treatment there were no significant differences between the groups. Before the treatments, the levels of oral inflammation were not correlated significantly with any of the serum or salivary immunoglobulin levels. However, the changes in oral inflammation were correlated significantly with the changes in the salivary IgM concentration after three and six months of treatment, and with the change in the salivary IgA concentration after six months of treatment.     

Colostral and serum IgG, IgA, and IgM concentrations in Standardbred mares and their foals at parturition

Immunoglobulin G, IgM, and IgA concentrations were measured in serum collected from 36 Standardbred mares within 12 hours of foaling, in colostrum collected within 6 hours of foaling, and in serum collected from foals 24 to 48 hours after birth. In serum collected from mares after parturition, mean concentrations of IgG, IgM, and IgA were 2,463.9 +/- 1,337.3 mg/dl, 136.4 +/- 218 mg/dl, and 305.2 +/- 237.5 mg/dl, respectively. In serum from foals, mean concentrations of IgG, IgM, and IgA were 1,953.3 +/- 1,635 mg/dl, 33.8 +/- 30.4 mg/dl, and 58.4 +/- 42.2 mg/dl, respectively. In colostrum, mean concentrations of IgG, IgM, and IgA were 8,911.9 +/- 6,282.2 mg/dl, 957 +/- 1088.1 mg/dl, and 122.9 +/- 77.3 mg/dl, respectively. The IgG concentrations in foal serum were poorly correlated with IgG concentrations in colostrum (r = 0.462, P less than 0.01). Correlations of IgM or IgA concentrations in serum from foals with IgM or IgA concentrations in colostrum and correlations of IgG concentrations in serum from mares with those in colostrum were not significant (P less than 0.01). Of 36 foals, 1 (2.8%) had a serum IgG concentration less than 400 mg/dl. Of 36 foals monitored for 4 months, 6 developed infectious respiratory tract disease requiring antimicrobial therapy at ages varying from 55 to 113 days; these infections were probably not related to failure or partial failure of passive transfer of antibody.     

Canine coronavirus infection in the dog following oronasal inoculation.

The pathogenesis of canine coronavirus (CCV) infection in 10-week-old puppies was studied up to 14 days after oronasal inoculation. Mild diarrhoea was seen from three to 11 days after inoculation, approximately coincident with faecal virus shedding. Virus was initially isolated from the tonsils on day 3, and then from both small and large intestinal tissues up to 14 days after inoculation. Virus was also isolated from liver and lung. Histological changes were not seen in any tissues, but CCV antigen was detected, using a peroxidase antiperoxidase staining technique, mainly in epithelium overlying gut-associated lymphoid tissue. Virus neutralising antibody was first detected on day 10. Specific anti-CCV IgM was first detected in plasma three days after inoculation and IgG on days 4 to 7. Small amounts of anti-CCV IgG, IgM and IgA were detected in duodenal secretion, but none in bile.     

Dynamics of Leishmania-specific immunoglobulin isotypes in dogs with clinical leishmaniasis before and after treatment

Concentrations of Leishmania-specific immunoglobulin G (IgG), immunoglobulin M (IgM), and immunoglobulin A (IgA) isotypes were analyzed by enzyme-linked immunosorbent assay (ELISA) in 23 dogs naturally infected with Leishmania infantum before and 1 year after initiating drug therapy. Results showed a high expression and prevalence of Leishmania-specific IgG (176.4 +/- 89 ELISA units [EU]), IgM (105.3 +/- 95.5 EU), and IgA (153.6 +/- 98 EU) in dogs before treatment (median +/- interquartile range EU). One year after treatment was started, dogs were classified as responsive dogs (RDs; n = 13) or unresponsive dogs (UDs; n = 10) based on clinicopathologic findings. Both groups of dogs experienced a statistically significant decrease (P < .05) in Leishmania-specific IgG (RDs = 27%, UDs = 41%), IgM (RDs = 42%, UDs = 29%), and IgA (RDs = 56%, UDs = 46%). Concentrations of specific IgG and IgM were not different at diagnosis or after treatment between the 2 groups. However, the median value for Leishmania-specific IgA 1 year after treatment was significantly lower (P < .05) in RDs (60.8 +/- 67 EU) than in UDs (117 +/- 54 EU). Examination of our data indicates that both the IgA isotype, which is mostly produced by mucosal plasma cells, and the IgM isotype are increased in infected symptomatic dogs, as previously reported for IgG. These 3 isotypes decreased significantly 1 year after initiation of medical treatment.     

Isotype-specific antibody responses to bovine coronavirus structural proteins in serum, feces, and mucosal secretions from experimentally challenge-exposed colostrum-deprived calves.

Five newborn isolation-reared colostrum-deprived calves were inoculated orally and intranasally when they were 20 to 30 hours old and challenge exposed when they were 21 days old with a suspension of virulent bovine coronavirus (BCV). Blood, feces, nasal swabs, tears, saliva, and bronchoalveolar lavage (BAL) fluids were collected from each calf prior to inoculation and then weekly for 5 post-inoculation weeks. An ELISA was used to quantitate the immunoglobulin isotype titers of BCV antibodies in all samples. An immunoblot assay was used to determine the antibody isotype responses to BCV structural proteins in all the samples, except saliva. At postinoculation days 2 to 3, all calves had severe watery diarrhea, shed BCV in their feces, and had evidence of BCV replication in their upper respiratory tract. After challenge exposure, no calves became ill and no evidence of BCV replication in the respiratory or intestinal tracts was detected. At postinoculation week 1, IgM responses to the N protein were seen in mucosal secretions (except nasal fluid) and feces. At postinoculation weeks 2 and 3, IgA was predominant in mucosal secretions and feces directed toward all the BCV proteins (except the E2 protein in BAL fluid). After challenge exposure, an increase (or failure to decrease) in most IgA and some IgG1 titers to BCV proteins was seen. The increases in IgA titers were to all viral proteins in all mucosal secretions and feces, except to the N and E1 viral proteins in feces. The IgG1 titer increases were to the E2 proteins in tears and BAL fluid and to the E3 protein in BAL fluid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation and phenotypic and functional characterization of cells from Peyer's patches in the dog.

Cells were isolated from canine Peyer's patches (PPs) and their phenotype and capacity to secrete immunoglobulins in vitro were determined. Cells isolated from duodenal and jejunal PPs of adult dogs consisted of 91.4% lymphocytes and 1.6% macrophages with 55.4% mIg(+)-cells and 35.6% Thy-1(+)-cells. In vitro IgA secretion by pokeweed mitogen (PWM)-stimulated PP cells exceeded that by cells from other lymphoid tissues and was specifically increased by concanavalin A, suggesting a role for isotype-specific T-cells. Comparison of duodenal and jejunal (proximal) PPs and the ileal PP revealed that the ileal PP contained fewer T-cells, fewer mIgA(+)-cells and more mIgM(+)-cells. Cells from the ileal PP produced very little IgA and IgG, but abundant IgM in vitro. These data suggest that the proximal PPs of dogs are important in the generation of IgA B-cells, similar to PPs of rodents. The ileal PP of dogs may have a function in the early development of the B-cell system of the dog.     

Immunoglobulin deficiency in Cavalier King Charles Spaniels with Pneumocystis pneumonia

Serum concentrations of immunoglobulin G (IgG), IgM, and IgA were measured in 9 Cavalier King Charles Spaniels with pneumonia caused by Pneumocystis sp that were examined at 4 veterinary surgeries in the United Kingdom (UK) between September 2001 and November 2002. Pneumocystis pneumonia was confirmed in all dogs by visualization of the organism in bronchoalveolar lavage fluid or a transthoracic lung aspirate. Two dogs had a history of demodicosis. Immunoglobulin concentrations also were measured in breed-and age-matched dogs sampled over the same period. IgG concentrations were significantly (P = .000) lower in the affected dogs (median 3.2 mg/mL) than in the control dogs (median 8.5 mg/mL). IgM concentrations were significantly (P = .002) higher in the affected dogs (median 1.95 mg/mL) than in the control dogs (median 1.12 mg/mL). One affected dog had no change in IgG concentration more than 3 months after resolution of infection or vaccination, but did have reduction in IgM concentration after resolution of infection and vaccination. Control dogs had low serum IgG and IgM concentrations, compared with the reference interval for all dogs. Lymphocyte count in blood was normal or high in 7 of 8 affected dogs. The results of this study suggest that there is a defect in immunity in Cavalier King Charles Spaniels that underlies the susceptibility of these dogs to pneumocystosis. Further studies are indicated to elucidate the mechanisms behind the defect, the prevalence within the breed, and the potential mode of inheritance of the problem.     

Hypogammaglobulinemia in Racing Alaskan Sled Dogs

Background: Serum immunoglobulin dynamics have not been studied in racing sled dogs, despite hypoglobulinemia having been reported during racing events.
Hypothesis/Objectives: Hypoglobulinemia in racing sled dogs is associated with decreases in serum IgA, IgE, IgG, and IgM concentrations during prolonged exercise.
Animals: One hundred and fifty-seven Alaskan sled dogs that successfully completed a 1,000 mile race.
Methods: Serum was obtained from 118 sled dogs within 1 month before the race and within 12 hours after completing the race. Serum also was obtained after 4 months of rest from 51 dogs that successfully completed the race, including 12 previously sampled dogs. Serum total protein ([TP]), albumin, and globulin ([Gl]) were measured, and serum IgA, IgE, IgG, and IgM were quantified by ELISA.
Results: The proportion of dogs with [Gl] ≤ 2.2 g/dL was significantly greater immediately after racing (38 of 118 dogs, 32.2%) than before racing (21 of 118 dogs, 17.8%, P = .005). Four months after racing, [Gl] was ≤ 2.2 g/dL in 23.5% (12 of 51) of dogs. [IgG] was significantly lower before (8.21 ± 4.95 mg/mL) and immediately after (7.97 ± 5.62) racing compared with 4 months after racing (18.88 ± 5.76). Serum [IgM] and [IgE] were higher and [IgA] was lower before racing compared with immediately after racing.
Conclusions and Clinical Importance: Sled dogs participating in long-distance racing have substantial decreases in [IgG] in addition to decreases in [IgM] and [IgE]. The pronounced hypogammaglobulinemia observed in a large proportion of racing sled dogs might predispose them to infectious disease.     

Immunological parameters in the intestinal lymph of pigs including changes during experimentally induced diarrhoea

Experiments were undertaken to provide basic immunological data on the intestinal lymph of young pigs. For this purpose indwelling cannulae were established in the main efferent intestinal lymphatic ducts of 12 animals and measurements were made of lymph flow rate, and concentrations of IgG, IgA and IgM. Measurements were also made of cell numbers, differential counts and immunoglobulin specificity of lymphoid cells in lymph. Similar measurements were also made on six pigs in which experimental diarrhoea was induced. The mean number of leucocytes in intestinal lymph was extremely low (0.66 X 10(5)/ml). However a high proportion of lymphocytes contained cytoplasmic IgA (19.65 per cent) and IgM (12.53 per cent), with few containing IgG (1.35 per cent). The concentrations of IgM and IgA in intestinal lymph were 0.51 mg/ml and 1.64 mg/ml respectively, values which suggest that the intestine is an important organ for synthesis of these two classes of immunoglobulin in young pigs. Following induction of diarrhoea and consequent dehydration of the intestine, the lymph: serum concentration ratios for immunoglobulins increased but subsequently declined when the water balance in the intestine returned to normal.