Mucosal and systemic isotype-specific antibody responses and protection in conventional pigs exposed to virulent or attenuated porcine epidemic diarrhoea virus

Eleven-day-old conventionally reared piglets were inoculated orally with two different doses of the cell-culture adapted strain CV-777 of the porcine epidemic diarrhoea virus (PEDV) or the virulent isolate of the same strain and challenged with the same virulent PEDV 3 weeks later. Pigs inoculated with the two doses of the attenuated virus did not show any typical sign of the disease, and virus shedding was not frequent. In contrast, 31% of pigs exposed to the virulent PEDV developed diarrhoea and virus shedding was demonstrated in 100%. At different postinoculation day (PID) and postchallenge day (PCD) virus-specific antibody-secreting cells (ASC) in gut associated lymphoid tissues (duodenum and ileum lamina propria and mesenteric lymph nodes) and systemic locations (blood and spleen) were assessed by enzyme-linked immunospot (ELISPOT). Only a small response was detected in the groups inoculated with attenuated PEDV, whereas in the group previously exposed to the virulent virus on PID 21 a large number of IgG and IgA ASC was detected. Isotype-specific antibody responses in serum were investigated by ELISA. IgG responses were detected in all groups, although the highest response corresponded to the group inoculated with virulent virus and only this group showed an IgA response. The pigs exposed to virulent PEDV were completely protected against the challenge with a higher dose of the same virulent virus on PID 21 and none of them shed the virus. The pigs inoculated with the attenuated strain were partially protected against the challenge, and 25% of the low dose- and 50% of the high dose-exposed pigs did not shed virus after challenge. All the pigs from a control group, not previously exposed to the virus, excreted the virus in faeces. A strong positive correlation was established between protection and the ASC responses detected in gut associated lymphoid tissues and blood at the challenge day and also between protection and serum isotype-specific antibody titers on that day. In addition, the IgA and IgG ASC responses detected in the blood on PID 21 also correlated with the responses found in the gut associated lymphoid tissues. The ASC and serum antibody responses after the challenge corresponded to a secondary immune response in the groups inoculated with attenuated virus, whereas a primary response was evident in the control group. No increase was seen in any of the parameters studied in the pigs inoculated with virulent PEDV.     

Helicobacter pylori-specific immunoglobulin synthesis in gnotobiotic piglets: evidence for the induction of mucosal immunity in the stomach

Short term tissue biopsy cultures and paired, sera, bile and gastric and intestinal contents from Helicobacter pylori-infected gnotobiotic piglets were tested for the synthesis of H. pylori-specific immunoglobulin (Ig) isotype production by antigen-specific ELISA from post-infection days (PIDs) 2-28. Serum antibody levels in all three Ig isotypes were elevated from baseline values by PID 14, serum IgM levels reached peak levels on PID 14 and by PID 28 bile was strongly positive for IgA and IgG.Intestinal, but not gastric contents from infected piglets, contained IgA-specific antibody from PID 14 onward. Gastric mucosal epithelia adjacent to areas of inflammation in infected but not uninfected control piglets produced readily detectable amounts of porcine secretory component (SC); IgA-positive plasma cells were identified in gastric submucosa and lamina propria in these areas. Culture fluid supernatants, collected from explanted gastric cardia and antra and intestinal ilea of H. pylori-infected piglets had trace amounts of IgA as early as PID 2 in some animals, and strong IgA reactivity in all by PID 28. Supernatants also contained H. pylori-specific IgG by PID 14. A strong gastric lymph node IgA response contrasted with moderate IgA production in mesenteric lymph nodes and spleen. Mucosal biopsy production of H. pylori-specific IgG was more evenly distributed throughout the lymphoid system. These data support the contention that the Ig response to H. pylori is initiated within the gastric compartment and matures over time to a generalized IgA-dominated mucosal and IgG-dominated nonmucosal humoral immune response.     

Modulation by colostrum-acquired maternal antibodies of systemic and mucosal antibody responses to rotavirus in calves experimentally challenged with bovine rotavirus

The effect of colostral maternal antibodies (Abs), acquired via colostrum, on passive protection and development of systemic and mucosal immune responses against rotavirus was evaluated in neonatal calves. Colostrum-deprived (CD) calves, or calves receiving one dose of pooled control colostrum (CC) or immune colostrum (IC), containing an IgG1 titer to bovine rotavirus (BRV) of 1:16,384 or 1:262,144, respectively, were orally inoculated with 105.5 FFU of IND (P[5]G6) BRV at 2 days of age. Calves were monitored daily for diarrhea, virus shedding and anti-BRV Abs in feces by ELISA. Anti-rotavirus Ab titers in serum were evaluated weekly by isotype-specific ELISA and virus neutralization (VN). At 21 days post-inoculation (dpi), all animals were euthanized and the number of anti-BRV antibody secreting cells (ASC) in intestinal and systemic lymphoid tissues were evaluated by ELISPOT. After colostrum intake, IC calves had significantly higher IgG1 serum titers (GMT=28,526) than CC (GMT=1195) or CD calves (GMT<4). After BRV inoculation, all animals became infected with a mean duration of virus shedding between 6 and 10 days. However, IC calves had significantly fewer days of diarrhea (0.8 days) compared to CD and CC calves (11 and 7 days, respectively). In both groups receiving colostrum there was a delay in the onset of diarrhea and virus shedding associated with IgG1 in feces. In serum and feces, CD and CC calves had peak anti-BRV IgM titers at 7 dpi, but IgA and IgG1 responses were significantly lower in CC calves. Antibody titers detected in serum and feces were associated with circulation of ASC of the same isotype in blood. The IC calves had only an IgM response in feces. At 21 dpi, anti-BRV ASC responses were observed in all analyzed tissues of the three groups, except bone marrow. The intestine was the main site of ASC response against BRV and highest IgA ASC numbers. There was an inverse relationship between passive IgG1 titers and magnitude of ASC responses, with fewer IgG1 ASC in CC calves and significantly lower ASC numbers of all isotypes in IC calves. Thus, passive anti-BRV IgG1 negatively affects active immune responses in a dose-dependent manner. In ileal Peyer's patches, IgM ASC predominated in calves receiving colostrum; IgG1 ASC predominated in CD calves. The presence in IC calves of IgG1 in feces in the absence of an IgG1 ASC response is consistent with the transfer of serum IgG1 back into the gut contributing to the protection of the intestinal mucosa.     

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.     

Specific antibody-containing cells in the mammary gland of non-lactating sheep after intraperitoneal and intramammary immunisation

Ewes were immunised intraperitoneally with ovalbumin and Brucella abortus in Freund's complete adjuvant, followed seven days later by intramammary immunisation in which ovalbumin was presented to one mammary gland and Brucella abortus to the other. Mammary tissue taken after a further seven days contained more antigen-specific plasma cells than ewes given intraperitoneal or intramammary immunisation alone. These cells were found predominantly in the specifically immunised gland and only a few were found in the contralateral gland. Most of these cells were of the IgG1 isotype. There was also an increase in the total number of IgG1- and IgG2-containing cells in mammary gland tissues of these ewes, indicating a non-specific response to immunisation. Following either intraperitoneal or intramammary immunisation there was also a significant increase in the number of antigen-specific IgA cells in the lamina propria of the jejunum. The gut response following intramammary immunisation alone was abrogated by chronic drainage of intestinal lymph but not mammary lymph. This suggests that antigen may relocate from the mammary gland to the intestine where an IgA response is generated from gut associated lymphoid tissue. These data provide evidence for interaction between the gut and mammary gland of sheep in response to antigen.     

A morphologic and immunohistochemical study of the bronchus-associated lymphoid tissue of pigs naturally infected with Mycoplasma hyopneumoniae

Porcine enzootic pneumonia (PEN), caused by Mycoplasma hyopneumoniae (Mh), has been described in pigs in all geographic areas. The disease is characterized by high morbidity and low mortality rates in intensive swine production systems. A morphologic and immunohistochemical study was done to determine the cellular populations present in lung parenchyma of infected pigs, with special attention to the bronchus-associated lymphoid tissue (BALT). Polyclonal and monoclonal antibodies were used for the detection of antigens of Mh, T lymphocytes (CD3+, CD4+, and CD8+), IgG+ or IgA+ lymphocytes, and cells containing lysozyme, S-100 protein, major histocompatibility complex class II antigen or myeloid-histiocyte antigen. Findings in lung tissues associated with Mh infection were catarrhal bronchointerstitial pneumonia, with infiltration of inflammatory cells in the lamina propria of bronchi and bronchioles and alveolar septa. Hyperplasia of mononuclear cells in the BALT areas was the most significant histologic change. The BALT showed a high morphologic and cellular organization. Macrophages and B lymphocytes were the main cellular components of germinal centers. T lymphocytes were primarily located in perifollicular areas of the BALT, lamina propria and within the airway epithelium, and plasma cells containing IgG or IgA at the periphery of the BALT, in the lamina propria of bronchi and bronchioles, in alveolar septa, and around bronchial submucosal glands. The hyperplastic BALT in PEN cases consisted of macrophages, dendritic cells, T and B lymphocytes, and IgG+ and IgA+ plasma cells. CD4+ cells predominated over CD8+ cells. Local humoral immunity appears to play an important role in the infection.     

Serum and mucosal antibody responses against Mycoplasma hyopneumoniae following intraperitoneal vaccination and challenge of pigs with M hyopneumoniae

Pigs were immunised intraperitoneally when six weeks old and again at about 10 weeks old with killed Mycoplasma hyopneumoniae antigen prepared in an oil adjuvant. The pigs were challenged with live M hyopneumoniae (Beaufort strain) at between 11 and 15 weeks old. Antigen specific antibody levels for both IgG and IgA classes in serum and respiratory tract secretion were monitored over time. In serum anti-M hyopneumoniae antibody was detected shortly after the second intraperitoneal vaccination and was largely IgG. In respiratory tract secretion the response was observed after challenge, and was primarily IgA. Anti-M hyopneumoniae antibody-containing cells and their immunoglobulin class specificity were monitored in lung and tracheal lamina propria. In lung the majority of anti-M hyopneumoniae-containing cells were IgG, whereas in the tracheal lamina propria the majority were IgA. These results are discussed in terms of the use of intraperitoneal vaccination for the control of M hyopneumoniae infection.     

Identification of porcine circovirus type 2 in retrospective cases of pigs naturally infected with porcine epidemic diarrhoea virus

The identification of porcine circovirus type 2 (PCV2) was studied in fresh intestinal tissues by polymerase chain reaction (PCR) and in formalin-fixed, paraffin-wax-embedded intestinal tissues by in situ hybridisation. The tissues came from pigs naturally infected with porcine epidemic diarrhoea virus (PEDV). A total of 35 (32.7%) of 107 small intestinal samples from pigs naturally infected with PEDV were found to be positive using PCR. Positive signals for PCV2 were detected in 32 (29.9%) of 107 small intestinal samples from pigs naturally infected with PEDV by in situ hybridisation. The distribution of positive cells in the jejunum and ileum was multifocal or patchy. Distinct positive labelling was found throughout the lamina propria in the small intestines. The results of this study indicate that PCV2 is highly prevalent in pigs naturally infected with PEDV.     

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.     

The Tissue Distribution of la- and IgM-Positive Cells in Adult and Newborn Miniature Pigs

Distribution of Ia- and IgM-positive cells was compared in tissues of adult and newborn miniature pigs, using indirect immunoperoxidase and immunofluorescent techniques with monoclonal antibodies. The different distribution patterns were found mainly in adult jejunum and kidney. Both IgM- and Ia-staining were ascertained in the lamina propria between the Lieberkühn crypts and in the lymphoid follicles of the jejunum. Moreover, IgM-positive cells were demonstrated in the crypts while Ia-reaction was seen in the lamina propria of the villi. In the kidney, IgM-staining was limited to the glomeruli. On the contrary, a distinct Ia-reaction was found mainly in intertubular structures. Small differences in IgM- and Ia-positive cell distribution were also noted in the mesenteric lymph nodes and in the spleen. Developmental changes of the adult distribution patterns (chiefly that of IgM-positive cells) were found in all tissues studied at 2 days of age.     

猪流行性腹泻免疫病理学研究Ⅳ.胃肠道IgA、IgM和IgG产生细胞数及其分泌液和血清中IgA、IgM和IgG含量动态观察

给10头8周龄仔猪经口感染猪流行性腹泻肠管毒,于感染后5、15、25、35和45d各扑杀2头,采取胃及各段肠管标本,用免疫过氧化物酶技术(间接法)检查胃肠粘膜固有层中IgA、IgM和IgG产生细胞数;收集血液及胃、各段肠管分泌液,应用ELISA双抗体夹心法测定IgA、IgM和IgG含量。结果:实验猪感染后第15d,空肠下段、回肠和回盲口处粘膜固有层中IgA和IgM产生细胞明显增多,肠管分泌液中IgA含量与IgA产生细胞数呈正相关,肠道局部免疫反应的高峰比全身性(系统性)免疫出现得早,且周期短。本试验提示,肠道粘膜积极参与了该病的免疫过程;胃肠道对猪流行性腹泻病毒(PEDV)免疫反应的主要部位在空肠下段、回肠和回盲目;参与胃肠道免疫反应的免疫球蛋白产生细胞主要是IgA和IgM产生细胞。     

Characterization and Distribution of B Cells in the Lymphoid Organs of Goats

The distribution of B cells in the lymphoid organs of the goat was studied using a panel of 13 monoclonal antibodies (mAbs) developed against different markers for bovine B cells. Samples of mesenteric lymph nodes, jejunal and ileal Peyer's patches, duodenum, jejunum, ileum, colon, caecum and rectum were taken from four 7-month-old male Murciano-granadina goats using the avidin-biotin-peroxidase (ABC) method on frozen sections as described by Hsu et al. (1981). The mAbs against immunoglobulins (Ig) recognized a large number of cells, particularly in the light zones of the germinative centres of the lymphoid follicles, regardless of the isotype against which they were directed. However, the greatest numbers of B cells in the germinative centres and outer coronas of the lymphoid follicles of the lymph node, spleen and Peyer's patches were recognized by mAbs against the L lambda chain of Ig and against IgM. This was also the case in other locations where B cells were abundant, such as the medulla of the lymph node and the dome of the Peyer's patches. These mAbs recognized not only B lymphocytes but also plasma cells, showing an intracytoplasmatic reaction (numerous in the spleen red pulp and the intestinal lamina propria when mAbs were used against the L lambda chain of the Ig, scarce in the intestinal lamina propria when used against IgM and scarce in spleen red pulp and numerous in the intestinal lamina propria when mAbs against IgA were used). The mAbs BAQ44 A, GC65 A and GB25 A are of interest because, besides marking cells in the B areas where lymphocytes show surface Ig, they give a positive reaction in areas where there are Ig-cells (the dark zone of the germinative centre) and do not immunostain plasma cells. Thus, these mAbs recognize a surface marker which is not an Ig.     

Immunohistochemical study of the local humoral immune system of the equine respiratory mucosa

An indirect immunoperoxidase technique was used to demonstrate both free immunoglobulin and immunoglobulin-containing plasma cells of IgG, IgA, and IgM classes in the mucosa of the equine respiratory tract. IgA-producing plasma cells predominated in the upper airways, whereas IgG-producing cells predominated in the lower respiratory tract. IgM-secreting cells were uncommon, but present in their highest numbers in the nasopharynx. Plasma cells specific for all of the immunoglobulin classes were identified in the surface epithelium, lamina propria connective tissue, glandular tissue and organised lymphoid tissue. The surface epithelium stained strongly for free IgA and IgM, but weakly for IgG, whereas glandular cells and the lamina propria connective tissue stained most strongly for IgG.     

Presence of immunoglobulins and antigens in serum, lung and small intestine in Ascaris suum infected and immunised pigs

The immunodetection of local Ascaris suum antigens and local and systemic antibodies were analysed in pigs reinfected with eggs or immunized with the 14, 42 and 97 kilodalton (kDa) fractions from A. suum. Twenty-one Iberian pigs were divided in 7 groups of 3 pigs. Groups 1 and 2 were uninfected and challenge control groups, respectively. Groups 3 and 4 were infected weekly with increasing doses of A. suum eggs and Group 4 was additionally treated with pyrantel pamoate. Groups 5, 6 and 7 were immunised with the 14, 42 or 97 kDa fractions from adult worms, respectively. Groups 2-7 were challenged with 10,000 infective eggs. Animals of Groups 3 and 4 showed a pulmonary granulomatous reaction with moderate number of eosinophils and leukocytes, while Groups 5-7 presented higher number of cells, especially in animals immunized with the 42 kDa fraction. These immunized groups presented abundant deposition of Ascaris body fluid (BF) and body wall (BW) antigens as well as the 14 and 42 kDa fractions in the pulmonary and intestinal tissues, while lower deposition of antigens was observed in animals of Groups 3 and 4. The immunized pigs of Groups 5 and 6 showed the highest systemic IgG titres in serum and these antibodies were negatively correlated with the number of larvae recovered in the lungs, suggesting that the IgG response may have a protective function against the ascariosis. The highest concentrations of IgA-bearing cells were observed in animals of Groups 3 and 4 compared to the immunised pigs (Groups 5-7), suggesting that local IgA production may be involved in the protection against migrating larvae. The main localisations of IgA-bearing cells were the bronchial and peribronchial areas of lungs and the lamina propia of duodenum. Low numbers of local IgG-bearing cells were observed in all animals and no IgM-bearing cells were detected in the local tissues.     

The differential localization of IgA,IgM and IgG in the gut of suckled neonatal piglets

The localization of immunoglobulins IgG, IgM and IgA in tissue sections prepared from the ileum of neonatal and adult swine were compared. Eighty percent of the immunoglobulin-containing lymphoid cells in the lamina propria of conventional adult German Landrasse swine were IgA-positive with lower numbers of IgM cells and occasionally an IgG cell. Anti-μ and α-chain reagents also stained the cytoplasm of the crypt epithelial cells. By comparison to these adult control tissues, the ileum of unsuckled neonates contained no immunoglobulins although after the ingestion of colostrum, the entire cytoplasm of the villus epithelial cells stained intensely when tested for IgG with only faint staining for IgM and IgA. On the other hand, IgA and IgM were readily localized on what appears to be only the apical border of the crypt epithelial cells but in contrast to the adult, the cytoplasm of these cells was unlabelled. IgG was absent from the crypt region. We interprete these findings to indicate an important, selective role for the villus epithelium in the absorption into the neonatal circulation of colostral IgG and probably IgA and IgM, and a specialized role for the crypt epithelium in adsorbing colostral IgA and IgM; possibly by complexing with mucin-bound secretory component.     

Detection of porcine epidemic diarrhea virus by immunohistochemistry with recombinant antibody produced in phages

Several diagnostic methods including immunofluorescence, enzyme-linked immunosorbent assay, polymerase chain reaction and immunohistochemistry have been developed for the detection of porcine epidemic diarrhea virus (PEDV). An immunohistochemical method using a new recombinant antibody produced by a phage antibody system (PAS16) kit was investigated and compared with that using a monoclonal antibody for PEDV detection in PEDV-infected piglets. In both the immunohistochemical methods, PEDV antigens were detected in the cytoplasm of villous enterocytes and in the macrophages infiltrated in the lamina propria at 18 to 110 hr post inoculation. The positive signals with the recombinant PAS16 antibody were similar to those with the monoclonal antibody. This result suggests that the recombinant PAS16 antibody can be applicable for the rapid immunohistochemical diagnosis of PEDV infection.     

Apoptosis in normal lymphoid organs from healthy normal, conventional pigs at different ages detected by TUNEL and cleaved caspase-3 immunohistochemistry in paraffin-embedded tissues

The frequency and the distribution of apoptotic cells were investigated in formalin-fixed paraffin-embedded lymphoid tissues from healthy conventional pigs at four different ages (6 days, 2 months, 3.5 months and 5 months). Samples of tonsil, mesenteric lymph node, spleen, thymus and Peyer's patches were histologically processed and apoptosis evaluated with the TUNEL reaction and cleaved caspase-3 immunohistochemistry. In each technique, quantification of positive labelling was done for each particular lymphoid tissue area. The labelling pattern and distribution were similar for TUNEL and cleaved caspase-3. TUNEL stained mainly apoptotic bodies inside macrophages, but signal was also seen in free apoptotic bodies and in the nuclei of lymphocyte-like cells. The anti-cleaved caspase-3 antibody labelled mainly nuclei of lymphocyte-like cells. All tissues presented a similar distribution pattern of apoptosis, except for the 6-day-old group. In this group, a scattered distribution of positive cells was detected in tonsil, lymph node and spleen. In the tonsil and mesenteric lymph nodes from the older pigs, follicular areas presented higher amounts of positive cells than interfollicular areas. Moreover, the splenic white pulp showed more positive reaction than the red pulp, especially when they included germinal centres. In all groups, the follicular areas of ileal Peyer's patches presented more labelled cells than the dome and the lamina propria. In the thymus, the higher apoptotic rates were found in the cortex. In general, TUNEL yielded higher rates of positive cells than cleaved caspase-3 immunolabelling. A good correlation between the two techniques was found for thymus, tonsil and mesenteric lymph node, but not for Peyer's patches and spleen. This study describes a detailed histochemical characterization of apoptosis in pig lymphoid tissues using TUNEL and a cleaved caspase-3 immunolabelling at different ages. Moreover, our results indicate that TUNEL and cleaved caspase-3 techniques can be equivalent only when tissues have a high or low levels of apoptosis, since a considerable discrepancy was found in intermediate situations. Data from this study should be useful for future comparative studies under disease conditions.     

Protective immunity following immunisation of pigs with aerosol of Actinobacillus pleuropneumoniae serotype 2

To introduce antigen to the respiratory mucosa, killed Actinobacillus pleuropneumoniae with quil A as adjuvant was administered to pigs as an aerosol. Immunisation by this aerosol induced a marked IgA response in the bronchoalveolar and nasal fluids, and in the serum. Following challenge with live bacteria two weeks after the last exposure to the aerosol, the immunised pigs were protected from the severe pleuropneumonia which developed in non-immunised pigs. The immunised pigs had lower antibody titres in the mucosal fluids and serum after exposure to the challenge. The immune response after experimental infection of non-immunised animals was a weak IgA antibody response in the bronchoalveolar and nasal fluids, whereas the systemic immune response after challenge included both IgA and IgG antibodies.     

Memory associated with IgA in the porcine respiratory tract

Pigs which had been immunised intraperitoneally with ovalbumin were subsequently challenged intratracheally with ovalbumin at intervals over a 200 day period. Intratracheal challenge 74 and 77 days after intraperitoneal priming resulted in a significant antigen-specific IgA response in respiratory tract secretion (RTS) on day 81. After a further 91 days, similar intratracheal challenge resulted in a lesser antigen specific IgA response in RTS. These trends were examined relative to antigen-specific IgA levels in serum and relative to antigen-specific IgG levels in serum and RTS. In addition, at the completion of the experiment the majority of anti-ovalbumin containing cells (AOCC) in the tracheal lamina propria were of the IgA class. Previously it has been shown that intratracheal challenge alone with ovalbumin had a negligible effect on AOCC in the trachea, and on specific antibody levels in respiratory tract secretion. The present findings suggest the existence of a memory mechanism operating in the porcine respiratory tract.     

Immunohistologic study of pulmonary and lymphatic tissues from gnotobiotic pigs inoculated with ara-T-resistant strain of pseudorabies virus

Ara-T-resistant strain of pseudorabies virus (PRV) was inoculated intranasally into six 2-week-old gnotobiotic pigs. Five inoculated pigs were sneezing and coughing. In pigs 1 to 4 killed on postinoculation days (PID) 3, 5, 7, and 9, respectively, PRV antigen was detected in respiratory epithelial cells, and pigs had severe pneumonitis. In pigs 5 and 6 killed on PID 11 and 13, respectively, PRV antigen was localized in macrophages in alveoli and necrotizing nodules. Immunoglobulin-containing cells (IgG, IgM, and IgA) were detected first in pneumonic lesions in pig 4 killed on PID 9. Detection of immunoglobulin-containing cells was coincident with pulmonary inflammation and regeneration of pneumonic lesions. The number of IgG-containing cells was greater than that of IgM- and IgA-containing cells. Corresponding to transient viral multiplication, IgG-, IgM-, and IgA-containing cells were demonstrated first in lymphatic tissues in pig 1 killed on PID 3 and their number was 5 to 10 times more than those in control pigs 7 and 8. Seemingly, PRV replication in lymphatic tissues stimulated the proliferative response of specific immunoglobulin-producing cells, and the appearance of immunoglobulin-containing cells in the lungs was associated with clearance of PRV and regeneration of pneumonic lesions.