Proteinuria and immunoglobulinuria in neonatal dogs

Samples of urine and serum from 45 newborn rottweiler puppies from six litters, and milk from their mothers, were taken 24, 48 and 72 hours and seven and 14 days after birth. Urine total protein and creatinine concentrations were determined and the ratios calculated. The immunoglobulin (Ig) concentrations of IgG, IgM and IgA in urine, serum and milk were determined with a commercially available elisa kit. The concentration of total protein in urine decreased from 1.64 to 0.29 mg/ml, and it and the ratio of total protein to creatinine in the urine of the neonatal puppies exceeded the normal values for adult dogs, but all the puppies developed normally. The average concentration of IgG in urine decreased from 0.0035 to 0.0003 mg/ml, that of IgA from 0.0035 to 0.0002 mg/ml and that of IgM from 0.0006 mg/ml to undetectable levels after two weeks. After two weeks, 47 per cent of the puppies had measurable levels of IgA and 70.6 per cent had measurable levels of IgG, but none of them had measurable levels of IgM.     

Porcine immunoglobulin transfer after prepartum treatment with selenium or vitamin E

Responses to prepartum injection of sows with Se and vitamin E (E) were evaluated by determining immunoglobulin (IgA, IgM, IgG) levels in the colostrum and serum of the sows and the serum of their offspring. Fifty-four sows (40 multiparous, 14 primiparous) receiving diets adequate in E and Se according to current NRC (1988) standards were randomly allotted to four treatment groups in which a single i.m. injection of saline (controls), 5 mg of Se, 1,000 IU of E, or both Se and E were given on d 100 of gestation. Sows were bled prior to and 7 d after injection, at farrowing and on d 14 and 28 of lactation. Colostral samples were collected at the initiation of farrowing. Pigs were bled 20 h postpartum and at 14 and 28 d of age. Major immunoglobulin changes in the serum of the sows due to treatment were not seen prior to parturition. Injections of Se and(or) E resulted in higher colostral IgM levels (8.4, 10.7, 9.8 and 9.6 mg/ml, respectively), but only the response from Se was significant (P less than .05). Concentrations of colostral IgA or IgG were not affected by treatment (P greater than .30). Compared with controls, all three treatments increased (P less than .10) IgM concentrations in serum from pigs at birth (28.3, 33.3, 36.0 and 33.5 mg/ml, respectively), whereas IgA and IgG concentrations were not affected (P greater than .30).(ABSTRACT TRUNCATED AT 250 WORDS)     

Maternal-neonatal immunoregulation: suppression of de novo synthesis of IgG and IgA, but not IgM, in neonatal pigs by bovine colostrum, is lost upon storage

Fifty-four neonatal pigs were allotted to 4 groups and reared in an electrically controlled automatic feeding device (autosow). Each group was reared on a different pool of bovine colostrum: fresh, stored 1 month, stored 6 months, and stored 8 years. Bovine and porcine immunoglobulins in the sera of these pigs, and in a group of conventionally reared pigs, were measured periodically during the first 42 days after birth. The maximal concentration of absorbed bovine immunoglobulin was reached between 12 and 18 hours and equaled or exceeded the amount of porcine immunoglobulin absorbed by the conventionally reared pigs. Large differences in the concentrations of the bovine immunoglobulin isotypes among the various pools of colostrum were positively correlated with concentration of these isotypes in the sera of the neonatal pigs fed these pools. Relative to their concentrations in colostrum, approximately 41% of the IgG1, 55% of the IgG2, 29% of the IgM, and 67% of the IgA was absorbed. The IgA was absorbed the best and IgM was least absorbed. Significant trends or differences in absorption were not observed among groups. Neonatal pigs given fresh colostrum, which had a higher fat content, had significantly more weight gain (P less than 0.05). This occurred, despite the fact that the fresh colostrum had the lowest concentration of bovine immunoglobulin. Serum half-lives for bovine IgG1 and IgG2 were significantly less than for porcine IgG (P less than 0.05), whereas the half-lives for bovine and porcine IgM and IgA were similar. De novo-synthesized immunoglobulins were detectable in serum after 6 days; IgM concentrations reached a maximum at 15 days in neonatal pigs given stored, but not fresh, colostrum.(ABSTRACT TRUNCATED AT 250 WORDS)     

The antibody-containing cell response in hepatic and intestinal lymph following intraperitoneal and intravenous administration of antigen in different adjuvants

A comparison was made of the antibody-containing cell (ACC) response in hepatic and intestinal lymph of sheep following intraperitoneal injection of ovalbumin in Freund's complete adjuvant (FCA), Freund's incomplete adjuvant (FIA) or the bland vegetable oil preparation, adjuvant 65. There was a substantial ACC response in intestinal lymph following intraperitoneal injection of ovalbumin in either FCA or FIA. Responses were essentially similar in magnitude and for both adjuvants ACC were distributed mainly among the IgG1, IgA and IgM immunoglobulin classes. In contrast, there was a negligible ACC response in intestinal lymph following injection of antigen in adjuvant 65. The output of ACC in hepatic lymph and the immunoglobulin class distribution of the ACC following intraperitoneal injection of antigen in either FCA or FIA were similar and results were comparable with those for intestinal lymph with ACC of IgA specificity comprising 32% of ACC at the peak of the response. In contrast to results for intestinal lymph, injection of antigen in adjuvant 65 was followed by a significant ACC response mainly of IgG₁-and IgM-specific cells in hepatic lymph. The ACC response was much smaller in magnitude than with the Freund's adjuvants. Intravenous injection of ovalbumin in FIA or adjuvant 65 gave rise to substantial ACC responses in hepatic lymph which contrasted with the barely detectable response in intestinal lymph. Following intravenous administration the great majority of ACC in hepatic lymph were of the IgM class irrespective of adjuvant used although ACC of the IgA class made a transitory appearance.     

Intestinal antibody response after vaccination and infection with rotavirus of calves fed colostrum with or without rotavirus antibody

The intestinal and systemic antibody response of calves vaccinated and/or challenged with rotavirus was studied employing isotype-specific ELISAs for the detection of IgG1, IgG2, IgM and IgA antibodies to rotavirus. Monoclonal antibodies to bovine immunoglobulin isotypes of proven specificity were used as conjugated or catching antibody. Five days after oral inoculation (dpi) of a 5-day-old gnotobiotic calf with rotavirus, IgM rotavirus antibodies were excreted in faeces, followed 5 days later by IgA rotavirus antibodies. The increase in IgM rotavirus antibody titre coincided with the inability to detect further rotavirus excretion. Faeces IgM and IgA rotavirus antibody titres fell to low levels within 3 weeks post infection. IgG1 and IgG2 rotavirus antibodies were not detected in faecal samples. In serum, antibodies to rotavirus of all four isotypes were detected, starting with IgM at 5 dpi. Two SPF-calves, which were fed colostrum free of rotavirus antibodies, were vaccinated with a modified live rotavirus vaccine and challenged with virulent rotavirus 6 days later. Upon vaccination, the calves showed an antibody response similar to the response of the infected gnotobiotic calf. Intestinal IgM rotavirus antibodies were excreted before or on the day of challenge and appeared to be associated with protection against challenge infection with virulent virus and rotavirus-induced diarrhoea. In 3 control calves, which were challenged only, the antibody patterns also resembled that of the gnotobiotic calf and again the appearance of IgM rotavirus antibodies coincided with the end of the rotavirus detection period. Two other groups of 3 SPF-calves were treated similarly, but the calves were fed colostrum with rotavirus antibodies during the first 48 h of life. These calves excreted passively acquired IgG1 and IgG2 rotavirus antibodies in their faeces from 2 to 6 days after birth. After vaccination, no IgM or IgA antibody activity in serum or faeces was detectable. Upon challenge, all calves developed diarrhoea and excreted rotavirus. Seven to 10 days after challenge low levels of IgM rotavirus antibody were detected for a short period. These data indicate that the intestinal antibody response of young calves to an enteric viral infection is associated with the excretion of IgM antibodies, immediately followed by IgA antibodies. This response is absent or diminished in calves with passively acquired specific antibodies which may explain the failure to induce a protective intestinal immune response by oral vaccination with modified live rotavirus of calves fed colostrum containing rotavirus antibodies.     

The distribution of immunoglobulin-containing cells in canine small intestine.

Peroxidase conjugated antisera to canine immunoglobulins G and A and to human immunoglobulin M, have been used to study the distribution of immunoglobulin-containing cells in the small intestine of the young and adult dog. At all levels of the small bowel of the adult dog, IgA:IgM:IgG cells have a 2:1:1 ratio while the greatest number of immunocytes is found in the duodenum and decreases towards the distal portion of the small intestine. The young puppy shows a predominance of IgM-containing cells in the small intestinal lamina propria during early life but the adult pattern of immunocyte distribution has been achieved by weaning.     

Antibody production by the pig colon during infection with Treponema hyodysenteriae

When 47 pigs were dosed orally with cultures of Treponema hyodysenteriae, 44 (94 per cent) developed swine dysentery. Of those which recovered and were rechallenged, nine of 21 (43 per cent) showed clinical signs, as did one of 10 (10 per cent) challenged on a third occasion. Clinical disease was associated with development of specific IgG, IgA and IgM antibodies in serum and the local production of IgA in gut mucosal tissues. The appearance of antibody was not directly related to protection but rather indicated either prolonged exposure (in the case of serum IgG) or recent exposure to T hyodysenteriae (for secretory IgA). Infection also resulted in the appearance of IgG and IgA memory cells in gut-associated lymphoid tissue. However, these studies indicated that humoral immunity alone is not responsible for the onset of a protective response to T hyodysenteriae in the colon.     

Dynamic changes of immunoglobulin concentrations in pig colostrum and serum around parturition

The aim of the study was the determination of IgA, IgM and IgG concentrations in porcine serum and colostrum, in order to evaluate their variations in the perinatal period, as well as to clarify whether there is a correlation between colostrum intake, initial level of immunoglobulins (Ig) in piglet serum and development of their own immunity. The mean IgA, IgM and IgG concentrations in sow serum 10 days before parturition were 1.58, 6.12 and 39.56 mg/ml, respectively. Seven days later only the IgG level was insignificantly lower (34.94 mg/ml, p = 0.55), while concentrations of IgA and IgM increased to 2.25 and 7.25 mg/ml, respectively (p = 0.23 and 0.62, respectively). The mean initial IgG concentration in colostrum at farrowing was 118.5 mg/ml and differed between sows. The average value of IgA in colostrum at birth was 23.8 mg/ml and decreased to 7.85 mg/ml at 6 hours (h) and to 4.59 mg/ml at 24 h after the onset of farrowing. IgM concentration at birth was 12.1 mg/ml and decreased to 4.23 mg/ml at 24 h postpartum. Positive relationships were found between concentrations of IgM and IgA in serum of piglets at 14 and 56 days of life (r = 0.41 and 0.80, respectively, p < or = 0.05) as well as for IgG concentration in the piglets serum at 7 days and 56 days of age (r = 0.48, p < or = 0.05). The above observations suggest that there is a correlation between the level of Ig in piglet serum in the first days of life and improvement of their own immunity.     

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.     

Relative deficiency in IgA production by duodenal explants from German shepherd dogs with small intestinal disease

Matched samples of serum, saliva and tears were collected from four groups of dogs; two of the groups were German shepherd dogs (GSDs) either with (Group 1) or without (Group 4) a variety of small intestinal disorders; the remaining two groups were dogs of other breeds, again with (Group 2) or without (Group 3) small intestinal disease. Capture ELISAs were used to measure IgG, IgM, IgA and albumin concentrations within these samples; intestinal humoral immune status of clinical cases was assessed by quantifying immunoglobulin production from duodenal explant cultures.There were no significant differences in IgG, IgM or IgA concentrations in serum, saliva or tears between the different groups of dog. Moreover, no significant differences were noted between groups for IgG, IgM and IgA salivary and tear secretory indices. IgA production by 24-h explant cultures was significantly lower in GSDs compared with non-GSDs with small intestinal disease (groups 1 and 2, respectively), but the numbers of lamina propria IgA(+) plasma cells in duodenal biopsies were not different between groups. These results suggest that there may be a relative deficiency in local IgA secretion in GSDs with small intestinal enteropathies, which is not reflected in either serum IgA concentrations, or in secretion at unaffected mucosal sites. It remains to be determined whether such a deficiency is a breed-related primary defect, or whether it arises secondary to the pathological processes within the intestinal mucosa.     

Computer model of the absorption and distribution of colostral immunoglobulins in the newborn calf

The transfer of immunoglobulin (Ig) isotypes (IgG1, IgG2, IgM), gamma-glutamyl transpeptidase (gamma-GT) and added D-xylose from colostrum to serum was investigated in newborn Holstein bull calves. Significant differences were observed in the time courses of the serum concentrations of these colostrum constituents following absorption from pooled colostrum. A computer model was devised to simulate the process of absorption of Ig isotypes, gamma-GT and D-xylose from colostrum in the newborn calf. A Fortran program was used to generate plots of the time course of the concentration of colostrum constituents in serum and other body fluids following a single feed of colostrum. These plots show how the changes in serum concentration of absorbed Ig isotypes, gamma-GT and D-xylose are affected by different rates of intestinal absorption, redistribution in body fluids and removal from plasma. A critical examination of data from the computer model and from the calf feeding experiments supports the view that the absorption of IgG1, IgG2 and IgM is not selective in the calf. The data were compared with earlier studies of the efficiency of the colostral transfer of Ig to the calf. In the present study the transfer efficiencies of IgG1, IgG2, IgM, gamma-GT and D-xylose were 46 per cent, 49 per cent, 47 per cent, 18 per cent and 21 per cent, respectively.     

Immunoglobulin isotype distribution of antinuclear antibodies in dogs with leishmaniasis

A modified indirect immunofluorescence method, using rat liver as substrate, was developed to determine the immunoglobulin isotypes forming antinuclear antibodies in sera from 12 antinuclear antibody-positive dogs out of 121 dogs with natural Leishmania infection. Immunoglobulin M was found to be the most frequent component of antinuclear antibodies (91·7 per cent), followed by IgG (41·7 per cent) and IgA (33·2 per cent). When these immunoglobulin isotypes were titrated, IgG antinuclear antibodies showed higher titres (1:200) than IgM and IgA antinuclear antibodies (1:50 and 1:20 respectively). Most of the antinuclear antibody-positive dogs simultaneously had two immunoglobulin isotypes, whereas none had all three immunoglobulin isotypes at the same time. Spearman rank correlation analysis showed no significant correlation between antinuclear antibody titres and circulating immune complexes or immunoglobulin levels. The low incidence of antinuclear antibodies and the absence of a clear relationship between isotype titres and clinical signs suggest a minor pathogenic role of antinuclear antibodies in canine leishmaniasis.     

Immunoglobulins in porcine umbilical cord blood and maternal placenta.

Studies were made of the immunoglobulin (Ig) in serums from umbilical cord of newborn pigs and maternal placenta. The neutralization test for porcine parvovirus and Japanese encephalitis virus was carried out with the serum of the sow and that of the umbilical cord of the newborn pig. Comparative studies of the serums from the dam and the umbilical cord were also done with gel filtration. Of 20 umbilical cord serum samples, IgG was seen in 5 samples (25%), IgA in 1 sample (5%), and IgM in 9 samples (45%). The amount of any 1 of the 3 classes of Ig in the serums was between 13.5 and 28.0 mg/dl. Among the samples examined, 1 had both IgG and IgA and 1 had IgG and IgM, but none had both IgA and IgM and none had 3 classes of immunoglobulins (i.e., IgG, IgA, and IgM). Only 7 samples (35%) did not have any class of Ig. The IgG disappeared from the blood of hysterectomy-produced colostrum-deprived pigs at 3 days of age, and IgM disappeared when pigs were 7 days of age. Neutralization antibodies of porcine parvovirus and Japanese encephalitis virus in maternal serum were not transferred to the fetus through the placenta. Results of immunohistologic surveys indicated that the sow's Ig were not transferred to the fetus through the placenta. Therefore, it is believed that the Ig in the porcine fetus might be synthesized in certain cells in the placental tissue, and the degree of production of the Ig in the placental tissues may differ in each case. The component, which seems to be Ig, was observed as the obscure band of the beta- to gamma-globulin area in serum of the umbilical cord. Comparison was made, with gel filtration, of maternal serum and serum from the umbilical cord of the newborn pig originating from the same sow. Seemingly, the IgG in the umbilical cord serum is mainly in the lower molecular weight fraction, whereas IgG in the sow's serum was distributed in the high to low molecular weight fractions.     

Antigen dose modulates the immunoglobulin isotype responses of pigs against intramuscularly administered F4-fimbriae

Parenteral immunisation normally induces a systemic antibody response characterised by high IgG and low IgA responses. In the present study, the effect of different doses of F4-fimbriae on the isotype-specific antibody response after intramuscular immunisation was studied in pigs. Pigs were injected twice with a 9 weeks interval with either 1, 0.1 or 0.01 mg of F4-ETEC fimbriae. The dose of 1mg F4 induced significantly lower primary F4-specific IgG and IgM responses than the doses of 0.1 and 0.01 mg F4, but primed for an enhanced F4-specific IgM serum antibody response after the booster immunisation. Furthermore, the dose of 0.1mg induced the highest F4-specific IgA serum response which was significantly higher than after injection with 0.01 and 1mg F4. Moreover, both lower doses (0.1 and 0.01 mg) showed a higher number of F4-specific IgA and IgG antibody secreting cells (ASC) in the local draining lymph nodes of the pigs. This study demonstrated that low doses of purified F4-ETEC fimbriae, especially the 0.1mg dose, are optimal for inducing F4-specific IgA responses after IM immunisation.     

IgA, igG1, IgG2, IgM, and BSA in serum and mammary secretion throughout lactation

Bovine IgG1, IgG2, IgA, and IgM were measured in the serum and lacteal secretions of six cows from 10 days prepartum to 240 days of lactation. Immunoglobulins in lacteal secretions were expressed in units of concentration (mg/ml) as well as in total daily output. All isotypes were selectively accumulated during colostrum formation. The rate of IgG1 accumulation decreased rapidly after calving; this decrease corresponded to a return to normal serum levels of this immunoglobulin. Selective accumulation of IgA > IgM > IgG1 was maintained throughout lactation, but IgG2 showed no selective accumulation beyond 5 days postpartum. In serum, IgA and IgM levels were elevated at parturition and showed a significant decrease postpartum. Increases in serum IgA levels 60 days postpartum corresponded to a rise in lacteal concentration. The concentration of all immunoglobulins increased during late lactation, coincident with a major reduction in milk yield. Six strains of mastitis-causing organisms were cultured during the period of the experiment; however, none resulted in clinical mastitis or showed an effect on immunoglobulin secretion.     

Quantitation of bovine immunoglobulins in culture fluids by use of sandwich radioimmunoassay with monoclonal antibodies

Bovine immunoglobulin isotype-specific murine monoclonal antibodies were used in sandwich radioimmunoassays to detect and quantitate bovine IgG1, IgG2, IgM, and IgA in culture fluids. The concentrations of bovine immunoglobulins in unknown samples were extrapolated from standard curves generated with bovine monoclonal immunoglobulins. The lowest detection limits for the bovine immunoglobulin isotypes ranged from 65 to 270 ng/ml.     

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.     

Normal profile of immunoglobulins in sera and tracheal washings of chickens

Concentration and distribution of the three immunoglobulins in the sera and tracheal washings of a chicken population was studied. The mean IgM, IgG and IgA concentrations in serum were 1.35, 5.09 and 0.31 mg/ml, respectively. The distribution of IgM and IgG in birds irrespective of age was almost normal whereas that of IgA was skewed. All the three immunoglobulins were present in tracheal washings but the level of IgM was barely detectable. The IgG was predominant in the tracheal washings but higher IgA : IgG ratio compared to that of serum indicated local IgA production in the chicken respiratory tract.     

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.     

Four-layer enzyme immunoassay (EIA) detection of differences in IgG, IgM and IgA antibody response to Aujeszky's disease virus in infected and vaccinated pigs

The use of the four-layer enzyme immunoassay (EIA) for the detection of IgG, IgM and IgA antibodies against Aujeszky's disease virus in blood and oropharyngeal swabs of infected and vaccinated pigs is described. Mean antibody titres obtained using the four-layer EIA were 6.1 and 3829 times higher compared with the indirect enzyme-linked immunosorbent assay (ELISA) and virus neutralization (VN) test, respectively. The VN test detected mainly IgG antibodies, while the IgM antibodies did not react. Using the EIA, the first antiviral antibodies in sera were demonstrated on Days 5-7 after infection or vaccination. Up to the 7th day, demonstrable antibodies were almost exclusively of the IgM class. In infected pigs high titres of IgM antibodies were still detected on Day 18, while in vaccinated animals they were absent by this time. Antibodies of the IgG class appeared in infected pigs sooner (Day 7) than in vaccinated pigs (Day 10) and reached higher mean titres. Antibodies of the IgA class were demonstrable from Day 10 only in samples from infected pigs. Similar antibody dynamics and distribution were detected in oropharyngeal swabs, except that the IgG and IgM titres were roughly 100 times lower than in sera. However, titres of IgA antibodies in oropharyngeal swabs were two times higher than in sera. The greatest differences between both groups of animals were recorded on Day 18; in the infected pigs, IgG, IgM and IgA antibodies were present in sera and oropharyngeal swabs at that time, while in vaccinated pigs only IgG antibodies were demonstrable. The effect of infection and vaccination on the pattern of the immune response as well as the importance of the detection of individual immunoglobulin classes for the specificity of the enzyme immunoassay are discussed.