Correlation between lymphocyte responses and immediate hypersensitivity to purified allergens

Seventy-nine unrelated subjects were selected for high allergic sensitivity to ragweed and/or grass pollens. Sensitivities to ragweed antigens E and Ra5 and rye grass group I were measured by intradermal skin testing and by histamine release in vitro. Lymphocyte responses were determined by incorporation of ³H-thymidine in vitro in the presence of antigen. Skin-test and histamine-release sensitivities were significantly correlated with lymphocyte response in the case of antigen E, but not for group I and Ra5. In addition, lymphocyte response to antigen E was significantly correlated with total serum IgE level. Six antigen E-sensitive individuals had marked delayed, but no immediate, reactions to Ra5 and almost all showed lymphocyte responses to Ra5. We conclude that immediate sensitivity and lymphocyte responses of allergic individuals to antigen E are significantly correlated, and that lymphocyte responses and delayed reaction to an antigen without immediate sensitivity or IgG antibody are possible even in allergic subjects.     

Localization of common (cross-reacting) antigens with Neisseria perflava and Klebsiella pneumoniae in tissues of the human bronchopulmonary apparatus

Antigenic similarity was studied between the microsomal fraction of tissues of the human bronchopulmonary apparatus and bacterial cells living in the respiratory tract:Neisseria perflava andKlebsiella pneumoniae. Cross reactions were studied with antimicrosomal sera in the complement fixation test withN. perflava andK. pneumoniae. Fixation of antibacterial antibodies and antibodies against microsomal fractions of the lung tissues was investigated in tissue sections of the human lungs and bronchi. The presence of antigens cross reacting with the antimicrobial sera was demonstrated in the microsomal fraction of tissues of the human bronchopulmonary apparatus.Allergologic Research Laboratory, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 2, pp. 210–212, February, 1976.     

Identification of IgG antibody as a carrier of reaginic activity in asthmatic patients

In a previous study it was reported that a group of asthmatic patients, in whom cromolyn sodium did not inhibit bronchial immediate allergic reactions, had reaginic antibodies that did not appear to belong to the IgE class. This study was designed to extend these observations, and it was shown that the IgG fractions from these patients' sera, purified by ion exchange chromatography and specific immunosorbents, had skin-sensitizing activity to the antigen studied, while the IgE fractions did not. The skin-sensitizing activity in the IgG fractions was not removed by anti-IgE antiserum and was not inactivated by heating at 56 °C or by reduction/alkylation. It is proposed that IgG antibodies may mediate immediate allergic reactions in some asthmatic patients, that these patients can be detected by means of serum total IgE and allergen-specific IgE estimation, and that cromolyn sodium is not an effective inhibitor of bronchial immediate allergic reactions in these patients.     

Immunochemical characterization of Aspergillus fumigatus antigens.

Culture filtrate antigens of Aspergillus fumigatus Ag 534 were purified by preparative isoelectric focusing and affinity chromatography. One of the pooled antigen fractions from the preparative isoelectric focusing step (pool 2) was passed through a concanavalin A column and yielded two components, designated antigens IIa and IIb. Antigen IIb reacted more strongly than antigen IIa with all of the aspergilloma and allergic bronchopulmonary aspergillosis sera tested by enzyme-linked immunosorbent assay. The glycoprotein nature of antigen IIb was shown by the concanavalin A binding properties and staining reactions of the components.     

Isolation, Characterization, and Immunogenicity of Mycoplasma pneumoniae Membranes

Membrane and soluble fractions of Mycoplasma pneumoniae, M. pulmonis, and M. laidlawii B were prepared by hypotonic lysis of whole cells. The membranes of M. pneumoniae and M. laidlawii B contained, as percentage of dry weight: 34 to 37% protein, 59 to 61% lipid, 3 to 4% carbohydrate as hexose, and 0.2% ribonucleic acid as ribose. NADH₂ and NADPH₂ oxidase activities were localized in the soluble fractions of M. pneumoniae and in the membrane fraction of M. laidlawii B. NADH₂ oxidase activity was localized in the soluble fraction of M. pulmonis. The lipids of M. pneumoniae were labeled when the organism was grown in the presence of either radioactive palmitic acid, oleic acid, cholesterol, or glycerol. The lipids were not labeled when grown in the presence of radioactive acetate. Palmitic acid radio-activity was found in neutral lipid, glycolipid, and phosphatide fractions. Immunodiffusion analyses of whole cells and membrane fractions demonstrated three reactive antigens. Two immunodiffusion antigens were localized in the membrane fraction. One of these apparently contains lipid. A third antigen, also considered lipoidal, was found in whole cells. Membrane and soluble fractions of M. pneumoniae were immunogenic. The immunogens eliciting metabolic-inhibiting antibodies were localized in the membrane. The membrane preparation also induced the formation of antibodies which fixed complement with an antigen extracted with lipid solvent. The soluble fraction contained a distinct immunogen which induces antibodies reactive in complement fixation with an antigen prepared by phenol extraction.     

Skin test reactivity to H. influenzae antigens as an outcome of the antigen structure and the balance between humoral and cell-mediated immunity in rats

Wistar RA rats were skin tested with several antigens prepared from Haemophilus influenzae, including a soluble (RE) and suspended form (CRE) of a somatic H. influenzae antigen, a preparation of bacterial capsular antigens (CA) and inactivated H. influenzae bacteria. The animals skin tested were passively sensitized with antigen-specific lymphoid cells or with antigen-specific antisera. Some animals were sensitized with a combination of specifically sensitized lymphoid cells and a specific antiserum.

Delayed hypersensitivity skin reactions were only observed after a cell transfer and using suspended antigen forms (CRE and inactivated H. influenzae). Using the soluble antigen form a delayed decreased reactivity was found after an identical cell transfer. On the other hand Arthus reactivity of the skin was only observed after a transfer of an antigen specific antiserum and using soluble antigen forms (RE and CA). The suspended form showed when used under these circumstances an opposite effect: a decrease of the normal inflammatory reactivity was found.

These results were interpreted as a support to the idea that the sensitization procedure (humoral vs cellular) is of importance to the character of the allergic reaction (immediate vs delayed); while the nature of the antigen seems to influence the intensity of the reaction (increase or decrease of the normal inflammatory response). Furthermore it was shown that antigen-specific antibodies were able to block the development of a d.h.-reaction to H. influenzae; while on the other hand specifically sensitized cells interfered with the development of an Arthus reaction to somatic H. influenzae antigen.

The usefulness of the described H. influenzae antigens as skin test antigens is probably dependent on the blocking effects of either humoral cellular immune systems and is discussed.

     

A further study of specific substances of the tularemia bacillus which induce a rapid allergic response

Summary It was found that fractions II and III (obtained by mild acid hydrolysis of the antigen of a virulentPasteurella tularensis culture) diluted 10,000 or more times could be used to test for the allergic response in subjects who hadhad tularemia. A specific allergic reaction was provoked within 15–30 minutes at the site of administration. A comparative study of the allergic response of guinea-pigs was investigated by means of the following preparation: antigens isolated from the bacterial mass of the virulent and vaccine strains (dried by acetone) by either Boiven's or Larson's method, an antigen isolated by Boiven's method from a suspension of living bacteria of the vaccine strain which had first been disintegrated by treatment with ultra-sound at a frequency of 1200 kc and 10 w/cm², and an antigen obtained from the same suspension by salting-out with 40% ammonium sulphate solution. The antigens isolated by the different methods from the virulent culture produced identical responses. The antigen from the vaccine strain produced a slower response, which was not pronounced until six hours had passed. It may, however, be used in human subjects in whom it causes an allergic reaction which is rapid in comparison with that elicited by tularin, and owing to the insignificant lipid fraction there are no side effects. Sterilization by autoclaving at one atmosphere pressure did not reduce the activity of the preparation. Disintegration of the bacterial suspension by ultra-sound reduced the allergenic activity of the antigen isolated from it.(Presented by Active Member AMN SSSR N. N. Zhukov-Verezhnikov) Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 52, No. 9, pp. 83–87, September, 1961     

Diagnostic accuracy of vaccine and vaccine excipient testing in the setting of allergic reactions to COVID-19 vaccines: A systematic review and meta-analysis

For persons with immediate allergic reactions to mRNA COVID-19 vaccines, skin testing (ST) to the vaccine/excipients (polyethylene glycol[PEG] and polysorbate 80 [PS]) has been recommended, but has unknown accuracy. To assess vaccine/excipient ST accuracy in predicting all-severity immediate allergic reactions upon re-vaccination, systematic review was performed searching Medline, EMBASE, Web of Science, and the WHO global coronavirus database (inception-Oct 4, 2021) for studies addressing immediate (≤4 h post-vaccination) all-severity allergic reactions to 2nd mRNA COVID-19 vaccination in persons with 1st dose immediate allergic reactions. Cases evaluating delayed reactions, change of vaccine platform, or revaccination without vaccine/excipient ST were excluded. Meta-analysis of diagnostic testing accuracy was performed using Bayesian methods. The GRADE approach evaluated certainty of the evidence, and QUADAS-2 assessed risk of bias. Among 20 studies of mRNA COVID-19 first dose vaccine reactions, 317 individuals underwent 578 ST to any one or combination of vaccine, PEG, or PS, and were re-vaccinated with the same vaccine. Test sensitivity for either mRNA vaccine was 0.2 (95%CrI 0.01–0.52) and specificity 0.97 (95%CrI 0.9–1). PEG test sensitivity was 0.02 (95%CrI 0.00–0.07) and specificity 0.99 (95%CrI 0.96–1). PS test sensitivity was 0.03 (95%CrI 0.00–0.0.11) and specificity 0.97 (95%CrI 0.91–1). Combined for use of any of the 3 testing agents, sensitivity was 0.03 (95%CrI 0.00–0.08) and specificity was 0.98 (95%CrI 0.95–1.00). Certainty of evidence was moderate. ST has low sensitivity but high specificity in predicting all-severity repeat immediate allergic reactions to the same agent, among persons with 1st dose immediate allergic reactions to mRNA COVID-19 vaccines. mRNA COVID-19 vaccine or excipient ST has limited risk assessment utility.     

Diagnosis of opisthorchiasis by enzyme-linked immunosorbent assay using partially purified antigens

Opisthorchis viverrini antigens were partially purified from adult worms collected from liver and extrahepatic biliary system of infected hamsters. Tegument fraction was obtained by chemical extraction, whereas other fractions were purified by Sephadex G-200 gel filtration chromatography. Five fractions of O. viverrini antigens were obtained, namely tegument extract, somatic extract, fraction 1 (P1), fraction 2 (P2) and fraction 3 (P3), respectively. The enzyme-linked immunosorbent assay technique was used to compare the reactivity of the five partially purified antigens. The sensitivity and specificity of all five antigens were compared by testing against the sera of 78 O. viverrini-infected individuals from O. viverrini endemic areas and 70 individuals from non-endemic areas infected with hookworm, Trichuris and Ascaris including 49 individuals with negative stool examination. The assays performed with tegument extract, somatic extract and P1 fraction were found to have 100% sensitivity, whereas the sensitivities of those with P2 and P3 were 96.1% and 83.3%, respectively. The tegument extract had the highest specificity as demonstrated by the lowest cross-reactivity with other parasites. Our results indicated that surface tegument is the most suitable antigen for use in immunological diagnosis of opisthorchiasis.     

Identification of Candida albicans antigens reactive with immunoglobulin E antibody of human sera.

Candida albicans antigens which reacted with immunoglobulin E (IgE) antibodies of 57 allergic patients were detected by immunoblotting. Of the various antigens, the 175-, 125-, 46-, 43-, and 37-kDa antigenic components reacted most frequently with the patient sera. To purify the major antigens, C. albicans cells were fractionated. The 46-, 43-, and 37-kDa antigens were recovered in cytoplasmic fractions, but the 175- and 125-kDa antigens were not recovered in any fraction. The 46-, 43-, and 37-kDa antigens were purified from cytoplasmic fractions by DEAE and P11 ion-exchange chromatography. Antigens were isolated by cutting bands out of sodium dodecyl sulfate-polyacrylamide gels. The purified components confirmed by immunoblotting were next processed for amino acid sequencing. Parts of the sequences of the 46-, 43-, and 37-kDa antigens had significant levels of homology with Saccharomyces cerevisiae glycolytic enzyme enolase, phosphoglycerate kinase, and aldolase, respectively. Rabbit IgG antibodies prepared against the 46- and 43-kDa antigens strongly cross-reacted with the homologous proteins of S. cerevisiae. However, S. cerevisiae enolase and phosphoglycerate kinase did not cross-react with IgE of patient sera. This result suggests that IgE antibodies against only small parts of their epitopes are elevated in the allergic patients. Since enolase is reported to be a major antigen for systemic candidiasis, this enzyme may be the immunodominant protein in both allergies and fungal infections.     

Type-III allergic reactivity toDiplococcus pneumoniae and encapsulatedHaemophilus influenzae in passively sensitized mice and rats

Haemophilus influenzae andDiplococcus pneumoniae are thought to play a role in the pathogenesis of chronic purulent bronchitis by producing type-III allergic reactions in the bronchial tree. It is also known that, depending on the size and form of the antigen, type-III allergic reactions may be produced that differ both at macroscopic and microscopic levels of observation.In this study we assessed the intensity and character of type-III allergic reactions produced by inactivatedD. pneumoniae, disintegrated pneumococci, and a crude soluble capsular substance prepared from this bacterial strain. Another set of bacterial antigens studied consisted of inactivated and encapsulatedH. influenzae, an aggregated and soluble form of a somaticH. influenzae antigen preparation, as well as a crude soluble capsular substance prepared from this bacterium.The bacteria and bacterial products were injected into the skin or into the peritoneal cavity of Wistar rats and C₃H mice. The experimental animals had been passively sensitized with specific antisera just before antigen injection.Our results confirm the hypothesis that the size and form of antigens strongly determine the character of type-III allergic reactions in animals with a given amount of circulating antibodies. Specific antibodies enhanced the intensity of reactions produced by disintegrated bacteria and soluble bacterial products, while on the other hand they were found to suppress reactions produced by intact, inactivated bacteria or aggregated bacterial products.In conclusion, when studying type-III allergic reactivity in chronic purulent bronchitis, one has to consider the size and form of bacterial antigens present in the bronchial tree.     

Antigens of mycobacterial cell walls

Three antigenic fractions from the cell walls of eight strains of mycobacteria were studied. Isolation and purification of these antigens were effected by enzymatic digestions, differential and sucrose gradient centrifugations, dialyses, and column chromatography. Two of the fractions were termed cell wall tuberculins (CWT-1, solubilized with lipase; CWT-2, solubilized with lysozyme); the third was termed “C” (cross-reacting) antigen. All appeared to be lipopolysaccharides. The CWT antigens, as compared with purified protein derivatives (human), were relatively species (group)-specific in both double immunodiffusion and guinea pig skin tests; in the latter, the reactions resembled those of delayed hypersensitivity. The C antigens reacted heterologously in double immunodiffusion and skin tests; the latter were the “immediate” type of reaction.     

Identification of Mycobacterium tuberculosis antigens in Seibert fractions by immunoblotting.

Seibert fractions prepared from Mycobacterium tuberculosis culture filtrates were evaluated by immunoblotting with a serum pool from patients with active pulmonary tuberculosis. Antibody activity was observed primarily with antigens in the polysaccharide II and A protein fractions; these fractions were further evaluated by immunoblotting with sera from individual patients with tuberculosis, from individuals without tuberculosis and positive for the purified protein derivative antigen skin test, and from individuals negative for the purified protein derivative antigen skin test. The antigens identified in the protein A fraction, a 32,000-molecular-weight antigen and a heterogeneous high-molecular-weight antigen, reacted with antibody found in sera from all patients with tuberculosis and with antibody from over 25% of the control individuals. A 10,000-molecular-weight antigen, a 30,000- to 44,000-molecular-weight antigen, and a heterogeneous high-molecular-weight antigen were observed in the polysaccharide II fraction; these antigens reacted with serum antibody from 70% or more of the patients with tuberculosis and with antibody from 20 to 70% of the control individuals. One of the antigens, with a molecular weight ranging from 17,000 to 28,000 in the polysaccharide II fraction, reacted with antibody in 64% of the sera from patients with tuberculosis but with only 1 of 15 control normal sera. This antigen may elicit an antibody response specifically associated with tuberculosis.     

Immunogenicity and specificity of collagen: V. Demonstration of three different antigenic determinants on calf collagen

Eighty rabbits were immunized with native or denatured acid-soluble calf skin collagen. Fifty-five antisera showed haemagglutination titres higher than 1:64. These sera were investigated with calf skin collagen, pepsin-treated calf skin collagen and rabbit skin collagen, used as coating antigens and inhibitors. Three serologically distinct collagen-antibody fractions could be demonstrated: a general, non-species specific fraction reacting with all three antigens, a species specific fraction active with the two calf preparations, and an antibody fraction to pepsin-labile structures of collagen reacting only with untreated calf collagen. The antibody fractions occur in the antisera either alone or as mixtures. The titres of distinct antibody fractions present in the mixed sera were determined by haemagglutination—inhibition experiments.

Antibodies to pepsin-labile structures could be equally well inhibited by collagen peptides obtained after trypsin or collagenase treatment. A ten-fold higher amount of peptides as compared with undergraded antigen was necessary to obtain the same inhibition effect. The results were compared with previously reported peptide inhibition experiments performed with the other two types of antibodies.

     

In Vitro Stimulation of Human Peripheral Blood Lymphocytes by Soluble and Membrane Fractions of Renografin-Purified Typhus Group Rickettsiae

Cell-free extracts of disrupted Renografin-purified Rickettsia typhi and R. prowazekii were evaluated as antigens in lymphocyte transformation assays for cell-mediated immunity to typhus group rickettsiae in 19 individuals with and 9 without histories of exposure to these organisms. Exposure consisted of clinical disease, vaccination with epidemic typhus vaccine, or occupational exposure to these agents. Both the soluble and membrane fractions of disrupted purified rickettsiae were used, and transformation of peripheral blood lymphocytes (PBL) was determined in microcultures by incorporation of [³]thymidine. Of the antigen concentrations tested (1 to 400 μg/ml), 10μg/ml appeared to be the most satisfactory. At this concentration, PBL transformation was highly reproducible and correlated well with donor exposure and the presence of enzyme-linked immunosorbent assay anti-typhus group immunoglobulin G. At higher concentrations, PBL from both exposed and control donors often responded to a lipopolysaccharide-like component present in these preparations. Specific transformation responses to rickettsial fractions were detected in several individuals decades after infection or vaccination, indicating that both fractions contained antigens associated with persisting cell-mediated immunity in humans. Generally, stimulation indexes with the soluble fraction were slightly greater than those obtained with corresponding concentrations of the membrane preparation, and in three individuals transformation was observed only with the soluble fraction. PBL transformation to soluble fractions also appeared to have some species specificity, since PBL from individuals with documented R. typhi infections were more responsive to the homologous soluble preparation than to the soluble fraction of R. prowazekii. PBL transformation also correlated well with homologous but only poorly with heterologous enzyme-linked immunosorbent assay immunoglobulin G titers.     

Immunological characterization of small nuclear ribonucleoproteins reactive with sera of patients with systemic lupus erythematosus

We have previously reported the purification of Sm and RNP antigens from goat liver and identified two polypeptides of molecular weights 70 and 80–90 kd as RNP specific and of 14 and 30 kd as Sm specific. In this communication the effect of ribonuclease and trypsin on Sm and RNP antigens was studied at the polypeptide level. We found that the RNP antigenic determinant polypeptides of 70 and 80–90 kd are lost as a result of such treatment, whereas there is no effect on the Sm-specific 14- and 30-kd polypeptides. The role of RNA in the antigenicity of Sm and RNP was studied by dissociation and reconstitution studies. The antigens were fractionated into protein and RNA and the individual fractions were tested for Sm and RNP activity by counterimmunoelectrophoresis (CIE) and enzyme-linked immunosorbent assay (ELISA). The RNA fraction did not react alone with anti-Sm and anti-RNP sera with either of the assays. Conversely when the protein fraction was tested by CIE, only Sm antigenicity was detectable. In the ELISA both Sm and RNP activities were demonstrated in the protein fraction. These results show that the presence of RNA is important in the immunoprecipitation reactions involving only RNP antigen, whereas Sm activity is independent of RNA. In addition, when the reaction is carried out by an assay involving primary antigen-antibody reaction (e.g., ELISA), RNP antibodies react with protein fractions alone, without the presence of RNA. We also report the glycoprotein nature of Sm-specific polypeptides. The antigen was found to react specifically with concanavalin A (Con A), indicating the presence of glycosyl and/or mannosyl residues. The observed glycoprotein nature of the Sm-specific polypeptides possibly explains their remarkable stability, unlike RNP-specific polypeptides, which are susceptible to proteolytic attack.     

Biochemical and immunological studies on soluble antigens ofEntamoeba histolytica

The soluble antigens ofEntamoeba histolytica trophozoites were analysed in detail by biochemical and immunochemical methods. The antigen was highly complex and heterogeneous as revcaled by Sephacryl S-300 column chromatography, which showed four distinct fractions. The molecular mass of fractions FI, FII, FIII and FIV was 660, 170, 65 and 13 kDa, respectively. Protein was the major constituent in crude soluble antigen (CSA) and fractions FI and FII (67%, 80% and 90%, respectively). Polysaccharide was predominant in the FIII fraction (59%). Antigenic activity observed after different physico-chemical treatments revealed that CSA and FI antigens were predominantly glycoprotein in nature. However, the antigenicity of FIII antigen was greatly reduced after sodium meta-periodate treatment, whereas no alteration in reactivity was discerned after trypsin treatment. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis demonstrated nearly 28 Coomassie blue bands for CSA and 20, 16, 15 and 3 polypeptide bands for the FI, FII, FIII and FIV fractions, respectively. The molecular mass of the polypeptides of these bands ranged from 210 to 20 kDa. Antigenic activity was observed in CSA and in the first three fractions, both in counter immunoelectrophoresis (CIEP) and in enzyme-linked immunosorbent assay (ELISA). However, the highest antigenic activity was noted in fraction FI. Major immunoreactive polypeptides of CSA and FI antigens against whole trophozoite antibody were observed in the 10- to 170-kDa regions. However, major differences in the immunoreactivity of the two antigens were noted at 116 and 14 kDa for FI antigen and at 84, 30 and 20 kDa for CSA. The binding of the FI antibody to the surface of the live parasite and the loss of immunoreactive polypeptides from the FI antibody after its adsorption with live trophozoites ofE. histolytica suggest a correspondence between FI and surface antigens ofE. histolytica. The efficacy of active immunization with CSA and its different fractions showed that antigenicity and immunogenicity were closely associated with the high-molecular-weight proteins, as 91% protection was observed for the FI antigens, whereas the FII and FIII fractions and CSA provided only 41%, 33% and 41% protection, respectively. These data suggest that FI antigen can be used in the serodiagnosis of and immunoprophylaxis against amoebiasis.     

Ribosomes of Acid-Fast Bacilli: Immunogenicity, Serology, and In Vitro Correlates of Delayed Hypersensitivity

Ribosomal fractions obtained from Mycobacterium bovis (BCG) and M. smegmatis (strain butyricum) were studied to determine their antigenicity, their ability to stimulate the production of soluble mediators of delayed hypersensitivity (in vitro correlates) by sensitized peritoneal exudate cells, and the antigenic relations of ribosomal antigens of BCG to BCG protoplasm and H37Rv culture filtrates. The crude ribosomes and the 50-30S ribosomal subunit pool obtained from each of the organisms induced both delayed and immediate hypersensitivity when injected in incomplete Freund adjuvant into rabbits, and skin reactions could be elicited in sensitized rabbits with those antigens. The crude ribosomes and 50-30S ribosomal subunit pool of M. smegmatis stimulated lymphocytes of guinea pigs sensitized with viable organisms to produce macrophage migration inhibition factor. Comparable ribosomal fractions from BCG bacilli caused lymphocytes of guinea pigs sensitized with viable M. bovis (BCG) to produce skin reactive factor. Immunoelectrophoretic studies showed that H37Rv culture filtrate, protoplasm, crude ribosomes, and 50-30S ribosomal subunits of BCG contain multiple precipitinogens and that many of these were shared between the different antigen systems. Comparative electrophoresis revealed that BCG protoplasm and H37Rv culture filtrate shared a major portion of their components with each other and relatively few with ribosomal systems. The ribosomal systems shared the major portion of their components with each other and relatively few with the other antigen systems.     

Experimental production of granulomatous pneumonitis: Comparison of immunological and morphological sequelae with particulate and soluble antigens administered via the respiratory route

Rabbits were sensitized with either a soluble protein antigen (BSA) or a particulate thermophilic actinomycete antigen (Micropolyspora faeni) via the respiratory route, followed by monitoring of sequential morphologic changes and the humoral plus cellular immunologic response. Primary respiratory tract sensitization with BSA resulted in a humoral anti-BSA response, Arthus and delayed skin reactivity, and in some cases specific antigen-induced alveolar macrophage migration inhibition, all in the absence of pulmonary lesions. Lesions characterized by mild multifocal perivascular mononuclear cell infiltrates in the lungs developed only after secondary BSA aerosol challenge. In contrast to these findings, “primary” respiratory tract sensitization with M. Faeni particulate antigen in saline solution resulted in the gradual development of extensive and progressive pulmonary interstitial and alveolar mononuclear cell infiltrates. These lesions were uniformly associated with specific serum precipitating antibody and delayed skin reactivity. Alveolar macrophage migration was significantly inhibited by Micropolyspora faeni in virtually of these animals. These results, while not excluding a primary irritant effect or Type II or III allergic tissue injury, suggest a role for delayed (cell-mediated) hypersensitivity in the pathogenesis of particulate actinomycete-induced pulmonary lesions. They also indicate that primary immunization with soluble purified protein antigens via the respiratory route can lead to systemic humoral and cell-mediated immunity without production of pulmonary lesions.     

Skin testing and extrinsic allergic alveolitis

Skin testing with six common allergens, tuberculin and a sterile avian antigen preparation from pigeon serum was performed on 102 pigeon fanciers. The incidence of positive prick tests to common allergens was no different for subjects with extrinsic allergic alveolitis. EAA, caused by avian exposure than the whole group. Positive immediate weal and flare reactions following skin prick testing with avian antigen occurred in 22 subjects and was closely correlated with atopy. However, when the same antigen was administered intradermally, 69 subjects developed an immediate (15 min) weal and flare reaction which did not correlate with atopy, instead, the weal diameter correlated significantly with the serum IgG antibody titre against pigeon serum gamma-globulin antigen, and furthermore, the higher grades of reaction were highly selective for subjects with EAA. Ten subjects, all with strong early intradermal skin reactions, developed a late (4-6 h) skin reaction; this was again highly selective for EAA. The subjects with cutaneous anergy to tuberculin had markedly higher IgG antibody titres to avian antigens, and these included the majority of the subjects with alveolitis.