Experimental infections with Campylobacter fetus in bulls of different ages

Factors affecting the establishment of a carrier state of Campylobactor fetus subsp. fetus (venerealis) in preputial cavities of bulls were investigated by following infection in bulls of two different age groups until slaughter, 9–18 weeks post infection. Each of the four older bulls (66–74 months at infection) and three of the four younger bulls (41–49 months at infection) were culturally positive until slaughter, indicating no increased susceptibility with age. Relative proportions of immunoglobulin classes and albumin in preputial fluids were generally similar to those determined prior to infection, although protein concentrations decreased and ratios of IgG/IgA and IgG₁/IgG₂ increased in most of the bulls. An unexplained diminution of sample volumes occurred with progressive sampling. Changes in concentrations of proteins and in sample volumes bore no apparent relationship to infection of bulls with C. fetus. Low levels of C. fetus agglutinins were detected in sample obtained both before and after infection, but no appreciable rise in antibody titers occurred following infection. Alterations in superficial antigens of C. fetus isolates obtained during the course of infection were demonstrated in the majority of animals. The capacity of the organism to undergo antigenic variation and to provoke a minimal immune response may contribute to its prolonged survival in the preputial cavity.     

Immunoglobulins and immunoglobulin-containing cells in the reproductive tract of normal rams

The levels of the immunoglobulins IgA, IgG1, IgG2, and IgM were measured in serum and fluid from various locations in the reproductive tract of normal rams. These fluids included semen, preputial washings, and fluid from the accessory sex glands (ASG), vasa deferens, rete testes, and tissue fluid from the seminal vesicles, bulbourethral glands, epididymal tails and efferent ducts. In addition, the prevalence of specific Ig-containing cells (ICC) was measured in sections of formalin fixed tissues stained by an indirect peroxidase-antiperoxidase labelling technique. Mean IgA levels in semen (1.23 mg/ml) and ASG fluid (0.46 mg/ml), were higher than in serum (0.19 mg/ml) and were at levels higher than IgG1 or IgG2 levels in semen, ASG fluid, and preputial washings, thus confirming the existence of a local immune system primarily in the ASG of ram genitalia. Relatively low concentrations of IgA and IgG in other genital fluids and IgG levels in these fluids were consistent with diffusion from serum. The relatively high prevalence of IgA-containing cells in bulbourethral (56% of all ICC) and prostate (49%) glands confirmed these tissues as major sites of local Ig production. ICC were also found in large numbers beneath pelvic urethral and preputial epithelia, but these were predominantly IgG-containing (88 and 72% respectively).     

Immunoglobulins and immunoglobulin-containing cells in the reproductive tracts of rams naturally infected with Brucella ovis

SUMMARY Thirteen rams with serological evidence of Brucella ovls exposure (CFT of 1:8 or greater), but with no or only mild epididymitis, were selected from a ram flock. Serum, semen, preputial washings and fluids from the accessory sex glands (ASGF) and testis and epididymis (TEF) were examined and immunoglobulin (lg) concentrations estimated. Genital tissues were examined histologically and the percentages of class specific immunoglobulin containing cells (ICC) determined. Eleven of these rams had histological evidence of active inflammation consistent with B. ovis infection; the organisam was cultured from the semen of 7. IgA concentration was high in semen (mean ± standard deviation of 5.03 ± 1.78 mg/ml) and ASGF (9.18 ± 7.28 mg/ml). These levels were much higher than those recorded in noninfected rams. IgA concentration was low in serum (0.78 ± 0.55 mg/ml) and TEF (0.59 ± 0.78 mg/ml). The concentrations of IgG¹, IgG², and IgM were low in all genital fluids sampled and not significantly different from those recorded in noninfected rams. This indicated that infection with B. ovis results in a pronounced IgA response in secretions, mostly from the accessory sex glands. Examinations of ICC, however, revealed that the plasma cell infiltrates of the epididymis, vas deferens, ampulla and seminal vesicle were predominantly IgG-containing (92.4, 97.2, 79.4 and 91.9% respectively). Fewer IgM-containing cells were scattered throughout these tissues, constituting 3.9, 6.3, 0.3 and 6.5% of all ICC, respectively. IgA-containing cells were most frequently seen in the ampulla (9.6% of ICC) where they were located directly beneath the epithelium, suggesting the ampulla as the most prominant location for the local production of IgA. These results indicate that although the tissue ICC response was predominantly IgG, IgA was selectively transferred into secretions with the exclusion of most IgG and IgM. ICC percentages in bulbourethral and prostate glands, and beneath pelvic urethral and preputial epithelia, were similar to those found in noninfected rams.     

Increased local IgA production in chronic obstructive pulmonary disease

The immunoglobulin (Ig) content of serum and tracheal lavage fluid was measured in 50 horses suffering from chronic obstructive pulmonary disease (COPD) and 40 control horses. The mean immunoglobulin: albumin ratios of the lavage fluids of both groups were significantly higher than the corresponding values for serum, which indicates significant local production of immunoglobulins in the lower respiratory tract. The IgA: albumin ratio of lavage fluid was significantly higher in diseased compared with normal horses, which implies increased local production of IgA in this disease. The IgG: albumin and IgM: albumin ratios of lavage fluid were not significantly different in the two groups of horses. These results reveal an involvement of the respiratory mucosal immune system in COPD.     

Changes in the concentrations of serum IgG, IgA and IgM of sows throughout the reproductive cycle

The concentrations of IgG, IgM and IgA in sera collected from 3855 sows (3208 pregnant and 647 lactating) at a single time point were determined. This experimental design allowed changes in serum immunoglobulin over the reproductive cycle to be studied without bias from seasonal influence. The concentrations of the three immunoglobulins changed independently during the reproductive cycle. Serum levels of IgM and IgG began a progressive postpartal decline during the 14th–17th week of gestation. At the onset of lactation serum IgG levels progressively increased while IgM levels continued to decline, the latter reaching their lowest level during the third week of lactation. In contrast to IgM and IgG, serum IgA levels increased 35% during weeks 14–17 of gestation and continued to increase throughout lactation, reaching their highest serum levels in the third week of lactation; the serum IgA concentration at this time was twice that observed during the first 13 weeks of gestation. Results of these studies allowed the reproductive cycle to be classified into four phases on the basis of serum immunoglobulin concentrations: (1) weeks 1–4 of gestation; (2) weeks 5–13 of gestation; (3) weeks 14–17 of gestation and (4) lactation.     

Quantification of immunoglobulins in respiratory tract secretions of the horse

Lavage techniques were used to obtain secretions from the nasal cavity, trachea and bronchi of conscious horses. The techniques, which utilised fibreoptic endoscopy for recovery of tracheal and bronchial secretions, were well tolerated by the horses. The recovery rates of the lavaged fluids were acceptable, but were lowest for bronchial secretions, and there was minimal contamination by blood. The fluids were analysed for IgG and IgM by single radial immunodiffusion, and for IgA and albumin by rocket immunoelectrophoresis. Relative to albumin there was significantly more IgA and IgM, and significantly less IgG, in the nasal cavity than the trachea. IgA and IgM levels were greatest in the nasal cavity and decreased progressively to the bronchi, whereas IgG levels showed the reverse trend. The immunoglobulin: albumin ratios of secretions taken from many levels of the tract were significantly higher than those of serum, suggesting local production of immunoglobulin in the respiratory tract.     

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.     

Demonstration of Tritrichomonas foetus in the external genitalia and of specific antibodies in preputial secretions of naturally infected bulls.

Portions of penis and prepuce were collected from 24 bulls with current or recent Tritrichomonas foetus infection. Epididymides were collected from seven of the bulls, and seminal vesicles and prostate were collected from four. Following immunohistochemical staining with two monoclonal antibodies (34.7C4.4 and TF1.15) prepared against T. foetus surface antigens, trichomonads were identified in sections from 15 of the bulls. Organisms were most often located in penile crypts in the midshaft and caudal regions and less often in preputial crypts. Trichomonads were not observed in sections from other genitalia or in subepithelial tissue. T. foetus antigen, however, was present in the cytoplasm of some epithelial cells and the cytoplasm of some mononuclear cells in subepithelial lymphoid aggregates and follicles. Preputial smegma was collected from 16 T. foetus-infected bulls and from 16 control bulls with negative T. foetus cultures. Preputial antibody levels to TF1.17, a surface antigen of T. foetus, were determined by an enzyme-linked immunosorbent assay. Preputial secretions from infected bulls contained specific antibody of each isotype and subisotype tested. IgG1 responses were the greatest, IgM and IgA responses were approximately equal, and IgG2 responses were low. Each isotype and subisotype response in infected bulls was significantly greater than that in the controls. These results confirm previous speculation concerning anatomical sites of infection and suggest that parasite antigen can be taken up and processed locally, resulting in deposition of specific IgG1, IgG2, IgA, and IgM antibodies in the preputial cavity.     

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.     

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.     

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.     

Immunoglobulin containing cells in normal and inflamed accessory sex glands of bulls

The peroxidase-antiperoxidase (PAP) technique was used to identify cytoplasmic immunoglobulins in the accessory sex glands of 15 normal bulls and 13 bulls with inflammation of the ASG. Immunoglobulin containing cells (ICC) of the types IgA, IgM, total IgG, IgG1 and IgG2 were measured and their percentages expressed. In accessory sex glands from normal bulls, IgA containing cells were the most frequent in prostate and bulbourethral glands (86.7% and 86.1%, respectively of all ICC present) whereas in the ampulla, IgG containing cells comprised 78.6% of the ICC. IgG1 and IgG2 containing cells were present in all the accessory sex glands in approximately equal numbers. Frequencies of IgM containing cells in the ampulla, prostate and bulbourethral glands were 6.3%, 4.0% and 3.7%, respectively. Although all isotypes of ICC were present in the seminal vesicle, the very low number precluded accurate quantification. In inflamed ampulla, seminal vesicle, bulbourethral gland and colliculus seminalis, IgG containing cells were the most frequent ICC with values of 66.2%, 83.0%, 69.0% and 53.5%, respectively; IgA containing cells were the second in prevalence with values of 21.5%, 10.3% 19.3% and 40.5%, respectively. The contribution of ICC to the locally protective immunoglobulins in accessory sex gland secretions is discussed.     

Allergen-specific IgG and IgA in serum and bronchoalveolar lavage fluid in a model of experimental feline asthma

Allergic asthma, a Th2 cell driven response to inhaled allergens, has classically been thought of as predominantly mediated by IgE antibodies. To investigate the role of other immunoglobulin classes (e.g., IgG and IgA) in the immunopathogenesis of allergic asthma, levels of these allergen-specific immunoglobulins were measured in serum and mucosal fluids. Bermuda grass allergen (BGA)-specific IgG and IgA ELISAs in serum and bronchoalveolar lavage fluid (BALF) were developed and optimized in an experimental model of BGA-induced feline asthma. Levels of BGA-specific IgG and IgA significantly increased over time in serum and BALF after allergen sensitization. Additionally, these elevated levels of BGA-specific IgG and IgA were seen in conjunction with the development of an asthmatic phenotype indicated by positive intradermal skin tests, enhanced airways hyperreactivity, and increased eosinophil percentages in the BALF.     

Immunoglobulins IgG and IgM in synovial fluids of swine with erysipelothrix polyarthritis

Concentrations of IgG and IgM immunoglobulins in synovial fluids and sera from a group of swine with experimentally produced Erysipelothrix rhusiopathiae polyarthritis were measured to determine if there was local synthesis of these immunoglobulins by plasma cells in arthritic synovial tissue. IgG and, to a lesser extent, IgM were significantly higher in arthritic than in nirmal synovial fluids from the same group of swine and this increase could only partly be explained by the increased permeability of the arthritic synovial membrane to plasma proteins. When synovial fluid values of IgG and IgM were calculated on the basis of companion serum concentration it was found that 82% of IgG, and 25% of IgM estimations were significantly elevated above levels in normal joints indicating that IgG was the dominant immunoglobulin synthesized.     

Local antibody responses in the bile and faeces of sheep infected with Fasciola hepatica

The effect of infection with the liver fluke Fasciola hepatica on serum, bile and faecal immunoglobulin and antibody levels was studied in Scottish Blackface sheep. In the serum the immunoglobulins showing the most marked increase were IgG1 and IgG2 and their maximal values were reached at 16 weeks after infection. In the bile IgG2 rose to peak values at two weeks and IgG1, IgA and IgM were maximal at four weeks after infection. The levels of faecal IgG and IgA were low after primary infection but after reinfection a rapid increase in IgA concentration was observed within one to two weeks. Haemagglutinating antibody levels against egg antigens, juvenile and adult excretory-secretory antigens and adult fluke somatic antigens were evaluated. In the sera high titres were observed starting from two to four weeks after infection and persisting until 14 to 16 weeks. Bile haemagglutinating antibodies against excretory-secretory antigens showed the highest level at two and four weeks after infection while antibodies against adult somatic antigens reached maximal titres between four and eight weeks. Faecal antibody levels after primary infection were low but increased rapidly within two weeks after reinfection, coinciding with the elevation in faecal IgA concentration. However, there was no reduction in the number of flukes established in reinfected animals.     

Immunoglobulins in normal porcine tracheobronchial secretions

Methods of collecting sputum and tracheobronchial washings are described.Proteins in normal porcine tracheobronchial secretions are studied by immunochemical qualitative and quantitative methods. Among the immunoglobulins, IgA, IgG and IgM are identified. Immunoelectrophoretic analysis of sputum shows albumin, transferrin and in the α-region 2 components not found in serum. The predominant immunoglobulin in tracheobronchial secretions is IgA, which accounted for about 85 % of the total immunoglobulin concentration in sputum and 61 % of the immunoglobulin concentration in tracheobronchial washings.     

Immunohistological studies of the local immune system in the reproductive tract of the mare

The immunoperoxidase technique was adapted for the identification of free immunoglobulin and immunoglobulin producing cells in equine tissues. Staining specific for free IgG, IgA and IgM was detected at all levels of the reproductive tract, and secretory component staining was present in the uterine epithelium but not in the oviduct, cervix or vagina. Immunoglobulin producing cells were present at all levels of the tract, with IgG and IgA cells at equivalent concentrations, but with fewer IgM cells. There was no cyclical trend in free immunoglobulin staining, or plasma cell numbers. IgG and IgM plasma cell numbers declined from uterus to vagina, as did epithelial staining, and the ratio of IgG:IgA cells declined from oviduct to vagina.     

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.     

Refractometry as a measure of the immunoglobulin status of the newborn dairy calf: comparison with the zinc sulfate turbidity test and single radial immunodiffusion.

Immunoglobulins were quantitated by single radial immunodiffusion in 34 female Holstein-Friesian calves which had been kept with their dams for the first 24 hours of life. The mean immunoglobulin G1 (IgG1) concentration was 1.063 g/dl; IgG2, 0.093 g/dl; IgM, 0.171 g/dl; and IgA, 0.125 g/dl. Both serum total protein as measured by refractometer and zinc sulfate turbidity measured at 498 nm gave good correlations with total protein, which were significant, P less than 0.001. Plasma total protein had a slightly poorer correlation with total immunoglobulins, presumably due to variable fibrinogen content. Plasma total protein gave a better correlation with total immunoglobulins than did any of the immunoglobulin classes individually. Total protein by refractometer underestimated naturally occurring or added immunoglobulins by one-third.     

Variations in the concentrations of the immunoglobulins IgG1, IgG2, IgM and IgA in sheep. 1. Changes in the blood serum and in the milk of sheep of different breeds and cross-breeds during lactation

A total of 103 ewes from the breeds Black mutt., Texel, Finn. L., Heidschnucke and the crossbreeds Texel x Finn. L. and Black. mutt. x Finn. L. was studied. Blood samples were drawn at days 1, 7, 21 and 42 and milk samples at the 4th, 12th, 24th and 72nd hour after the onset of lactation and subsequently on days 7, 21 and 42. The concentrations of the immunoglobulins IgG1, IgG2, IgM and IgA were assayed in serum and milk. The following results were obtained: 1. The total immunoglobulin contents in the serum was not significantly different between breeds. 2. All ewes showed a rise in serum immunoglobulin concentrations by about one third over the first six weeks of lactation. Between 50-60% of this increase were on the account of IgG1. 3. The serum concentration of IgG1 and IgG1 rose as of the third day, those of IgM as of day 21 after lambing. 4. The rise in serum immunoglobulin concentration continued after the weaning of the lambs. 5. The ratio of IgG1 to IgG1 in ewe serum was 2:1. 6. The immunoglobulin concentration in milk dropped sharply on the first day of lactation, followed by a continuous, more gradual decrease over the entire course of lactation. A terminal rise, as observed in sows, could not be detected. 7. The ratio of IgG1:IgG2 : IgM : IgA in the whey changed from 85 : 1 : 12 : 2 on day one to 70 : 7 : 12 : 11 on the last day of lactation. 8. While characteristic trends in immunoglobulin patterns in the sera of ewes over the course of lactation are clearly discernible, it is not possible to denote "normal" values.