New developments in immunopathology with special reference to complement and immune complexes

The last few years have seen rapid advances in our understanding of immune phenomena, and these are now being translated into a new understanding of immunopathology. This paper reviews a number of areas where important new knowledge has been gained in the area of complement and immune complexes as a cause of disease. An understanding of the role of the metabolism of complement proteins has led to an understanding of how these proteins interact with receptors to cause phagocytosis. New understanding of the lysis-producing steps has led to new diagnostic tests for complement-induced damage as well as new knowledge of how complement functions to kill microorganisms. New methods for purifying anaphylatoxins have made these proteins available for study in man. A model of serum sickness in man has allowed for the more detailed understanding of how immune complexes cause disease and has led to the appreciation of new clinical signs of immune-complex-mediated disease in man.     

The fate of vacuolation in liver cells with special reference to hyaline globules.

This study was undertaken in rats to clarify the mechanisms and time necessary for recovery from vacuolation in liver cells. Vacuoles were produced by congestion of the liver due to constriction of the inferior vena cava just below the diaphragm, and changes in vacuoles were examined quantitatively and qualitatively until 24 h after release of the constriction. Vacuoles in liver cells decreased in number by half within 5 min after recovery from congestion. The remaining vacuoles metamorphosed to hyaline globules by condensation of the contents. The number of hyaline globules increased with a peak occurring at 3-6 h after recovery from congestion, although the number of vacuoles decreased gradually. Only a few small vacuoles and hyaline globules were found in liver cells in pericentral areas at 24 h after recovery from congestion. These data indicate that vacuoles may be discharged promptly from the liver cell cytoplasm after recovery from congestion, and the remaining vacuoles may metamorphose to hyaline globules by condensation of the contents and finally fade into the cytoplasm.     

Patient education for children with interstitial lung diseases and their caregivers: A pilot study

ObjectivePatient education in children with rare chronic diseases like children’s interstitial lung disease (chILD) remains a challenge.AimsTo develop and evaluate a component-based educational program for individual counselling and to improve patients’ and caregivers’ self-efficacy and treatment satisfaction. Furthermore, to create chILD-specific educational material and assess physicians’ satisfaction with the intervention as well as patients’ health-related quality of life (HrQoL).MethodsThe study was conducted in two German centers for pediatric pulmonology, as a single-group intervention with pre-post-follow-up design.ResultsParticipants (N = 107, age: M = 7.67, SD = 5.90) showed significant improvement of self-efficacy (self-report: t = 2.89, p < 0.01; proxy-report: t = 3.03, p < 0.01), and satisfaction (patients: t = 3.56, p = 0.001; parents t = 6.38, p < 0.001) with the medical consultations. There were no pre-post differences in HrQoL. Participants were highly satisfied with the material and the physicians with the program.ConclusionsThe chILD education-program is a promising strategy to improve patients’ and their parents’ self-efficacy and treatment-satisfaction. Specific effects of the intervention need to be determined in a randomized controlled trial.Practice implicationHealthcare providers managing pediatric patients with chILD, may choose to use a patient education-program specifically tailored to the needs of chILD patients and their families, such as the program described here, which is the first of its kind.     

Cell-specific regulation of gamma-glutamylcysteine synthetase in human interstitial lung diseases

The pathogenesis of interstitial lung diseases (ILDs) is known to be associated with reactive oxygen and nitrogen metabolites and increased oxidant stress. One of the major antioxidants in human lung is glutathione (GSH) and enzymes linked to its synthesis. The rate-limiting enzyme of GSH synthesis is gamma-glutamylcysteine synthetase (γ-GCS) containing catalytically active heavy (γ-GCSh) and regulatory light (γ-GCSl) subunits. It can be hypothesized that γ-GCS is the major determinant in explaining reduced GSH levels in fibrotic lung disorders. We investigated the regulation of γ-GCS by transforming growth factor beta₁ (TGF-β₁) and tumor necrosis factor alpha (TNF-α) in human lung cells and its expression and distribution in fibrotic (biopsy-proven idiopathic pulmonary fibrosis, for instance, usual interstitial pneumonia, UIP, n = 15), inflammatory, and granulomatous diseases of human lung parenchyma (desquamative interstitial pneumonia, n = 10; ILD associated with collagen diseases, n = 10; sarcoidosis, n = 19 and allergic alveolitis, n = 8). In human lung alveolar epithelial cells, γ-GCSh was decreased by TGF-β₁, whereas TNF-α caused a transient enzyme induction. In normal lung, γ-GCS was mainly localized to the bronchiolar epithelium. In UIP, the highest immunoreactivities were observed in the airway epithelium and metaplastic alveolar epithelium, but fibroblastic foci were negative. In sarcoidosis, the highest reactivities were detected in the epithelium, alveolar macrophages and pulmonary granulomas. γ-GCS was ultrastructurally localized to the cytoplasm of regenerating type II pneumocytes and macrophages. In conclusion, γ-GCS is widely expressed in sarcoidosis and regenerating epithelium but is low in the fibrotic areas of usual interstitial pneumonia, probably because of enzyme down-regulation.     

Local immune complexes and inflammatory response in patients with chronic interstitial pulmonary disorders associated with collagen vascular diseases

Evidence is accumulating that the lung injury in collagen vascular diseases (CVD) is triggered by immune complexes (IC). These reactions are neutrophil- and complement-dependent. The direct, in vivo phagocytosis of IC by bronchoalveolar lavage polymorphonuclear leucocytes (BAL-PMN), was studied in 15 patients with CVD and chronic interstitial pulmonary disorders. A control group (NC) consisted of nine healthy, non-smoking volunteers. Concentrations of soluble IC were measured using a solid phase Clq ELISA assay, and an indirect, in vitro phagocytosis assay performed using healthy donor PMN. Local Ig and C3 concentrations were determined using laser nephelometry and Mancini techniques, respectively. In the patient group the total cell counts/ml recovered lavage fluid and the proportions of BAL-PMN were significantly increased (P less than 0.05 and P less than 0.001, respectively). The influxed PMN showed high scores of direct IC phagocytosis. Soluble IC concentrations were significantly increased compared with controls (all comparisons P less than 0.01), and in the BAL relatively higher than in the serum of the same patients. Concomitantly high local IgG concentrations were observed. Corticosteroid treatment gave rise to significantly decreased total cell counts (P less than 0.05), and proportions of BAL-PMN (P less than 0.001), a decrease in the in vivo IC phagocytosis (P less than 0.05), in the indirect, in vitro IC phagocytosis, in the Clq ELISA and in the local IgG concentrations (all comparisons P less than 0.001). We concluded that locally formed IC may induce an inflammatory response in the lungs of patients with CVD.     

Studies on NEM-treated erythrocyte clearance in the rabbit, with special reference to the effects of circulating immune complexes.

The splenic component of mononuclear phagocytic cell function was investigated in rabbits using as a marker the clearance of N-ethylmaleimide-treated, technetium-labelled autologous erythrocytes. In 66 clearances performed in 23 normal rabbits, the clearance T1/2 was found to be between 5 and 16 min, the value for each rabbit remaining relatively constant. Clearance was proportional to incubation time or the dose of N-ethylmaleimide used. Clearance was delayed following injection of 20 to 65 mg of preformed complexes of BSA-anti-BSA made in 10-fold antigen excess. The degree and duration of this blockade was related to the dose of the complexes up to a critical value of 20 mg (antibody content after which no further decrease in clearance occurred although the duration of the blockade was longer at higher doses. Ultracentrifugation studies showed that these complexes were about 14S in size and sequential studies revealed that they were slowly cleared from the circulation. The experiments indicate that studies of splenic function may be of value in assessing immune complex disease, since circulating immune complexes of these characteristics are poorly detected by current methodology.     

Immunohistochemical characterisation of pulmonary hyaline membrane in various types of interstitial pneumonia

AIMS: Hyaline membrane (HM) in diffuse alveolar damage (DAD) pattern is frequently detected in the acute stage of interstitial pneumonia (IP). To determine the exact nature of HM, we investigated immunohistochemically 25 cases of HM-containing IP. METHODS: The cases examined using various kinds of antibodies were four cases associated with rheumatoid arthritis, five with usual interstitial pneumonia, two with dermatomyositis, five with viral infection, one case with progressive systemic sclerosis and eight cases caused by other agents. RESULTS: HM mostly reacted with antibodies to PE10 (SP-A), Factor VIII, KL-6 and EMA and, interestingly, stained for AE1/AE3, CK19, and Hup-1 in some cases, but was negative for PTAH staining. However, the immunoreactivities of HM varied even within the same disease or section. CONCLUSIONS: The immunohistochemical heterogeneity of HM suggests that HM may be formed by different mechanisms in various types of IP. Our findings also suggest that the main components of HM are derived from alveolar epithelial cells and their proteins, some including cytoplasmic element of CK19, and also from serum factors, but not fibrin. The immunohistochemical characteristics of HM in DAD pattern will aid understanding of the significance of HM formation in IP.     

A histopathological and ultrastructural study of eccrine porocarcinoma with special reference to its subtypes

Summary Five cases of eccrine porocarcinoma were studied by light and electron microscopy. Histopathologically, these could be classified into two types; the common and the giant cell type. The common type was characterized by almost uniform medium-sized cuboidal tumour cells and a formation of well-developed intracytoplasmic lumina. A broad diversity of histopathological and ultrastructural features was seen in these tumours. The tumours of the giant cell type consisted of mononu-clear polygonal cells and bizarre giant cells. This type was considered to be an undifferentiated form of porocarcinoma.     

A five year (1985-1989) retrospective study of equine neurological diseases with special reference to rabies.

A retrospective study of horses necropsied between 1985 and 1989 at a diagnostic laboratory of a veterinary school in North America is documented. In this investigation over 20 per cent of the horses had clinical neurological signs. Equine protozoal myeloencephalitis (caused by Sarcocystis neurona) and cervical stenotic myelopathy (wobbler syndrome) were the most common of these disorders. The veterinary school is located in the midst of a raccoon rabies enzootic area. However, only four cases of equine rabies were diagnosed during the 5-year study. The gross microscopical and immunohistochemical findings from these rabies-positive horses are documented. Immunoperoxidase tests for detection of rabies antigen in another 35 horses with non-specific encephalitis/encephalopathy did not reveal any positive cases. Based on this investigation, it appears that immunoperoxidase is a valid method for diagnosis of rabies when fresh tissues are not available for the fluorescent antibody test. It is also concluded that no cases of equine rabies were overlooked by the diagnostic laboratory during the period under investigation.     

A simple method to quantitate circulating immune complexes in different diseases

Immune Complexes are involved in the Pathogenesis of many diseases of varied aetiology such as autoimmune disorders, protozoal diseases, bacterial and viral infections. Quantitation of immune Complexes in these diseases can be used for diagnosis and to ascertain the prognosis. The simple method of precipitation by polyethylene glycol and quantitation by single Radial Immunodiffusion has been used in leprosy, syphilis, bacterial endocarditis and systemic lupus erythematosus (SLE). This method found significantly higher levels of circulating immune complexes (CICs) in erythema nodosum leprosum, culture positive bacterial endocarditis and SLE where CICs are known to play an important role in the pathogenesis.     

Immunoelectron microscopic study of S-100 beta-positive human T-lymphocytes with special reference to double immunostaining

S-100 beta-positive T-lymphocytes in human peripheral blood were shown to be definitely T-cells containing the S-100 beta subunit in their cytoplasm by double immunostaining using immunoferritin labeling for CD8 antigen and immunoperoxidase labeling for cytoplasmic S-100 beta subunit at the ultrastructural level. These results ultrastructurally confirmed the presence of double markers of S-100 beta-positive T-lymphocytes seen by light microscopic double immunostaining. Furthermore, this double immunostaining method for electron microscopy is applicable to other kinds of cells for the simultaneous detection of two different antigens present both on the cell surface and in the cytoplasm.     

Enzyme immunoassay detection of circulating immune complexes by monoclonal antibodies to C3 neoepitopes with special reference to IgG concentration and to interfering anti-immunoglobulin antibodies

A sensitive solid-phase anti-C3 enzyme immunoassay for detection of circulating immune complexes (CIC) is described. A mixture of the monoclonal antibodies (MoAbs) bH6 and Clone 9 specific for neoepitopes on C3 activation products was used as capture reagent. MoAb bH6 recognized C3b, iC3b and C3c, and Clone 9 recognized iC3b and C3dg. Detection antibody was a polyclonal peroxidase-conjugated rabbit anti-human Ig antiserum. A quantitative assay was constructed using serum incubated with heat aggregated IgG (HAG) as standard. The lower detection limit was 5 micrograms/ml of HAG. Interassay and intra-assay coefficient of variation was 15% and 5%, respectively. Anti-animal immunoglobulin antibodies were detected both in normal and pathological sera. This activity was efficiently absorbed by nonimmune immunoglobulins added to the samples. The present assay was compared with a polyethylene glycol precipitation assay for CIC determination. The latter assay was strongly influenced by the IgG concentration (rs = 0.78; P = 0.006), whereas no such correlation was seen for the anti-C3 immune complex assay (rs = -0.30; P = 0.20).     

A study of cancer of the breast with special reference to its causation and prevention

Breast feeding, lactational histories, parity, age at marriage, and socio-economic status were compared in 24 patients with carcinoma of breast, 24 healthy controls, and 48 patients suffering from other diseases. They were matched for age, social class and work or trade of the husbands. Breast cancer patients married later, had shorter lactational histories and had fewer children as compared with controls. Studies in six healthy mothers showed that milk became more alkaline on stasis in the breast. This study confirms the view that breast feeding protects against breast cancer. It suggests that one carcinogenic factor may be an alkaline milieu produced by the stasis of milk in the breasts. An alkaline milieu surrounding epithelial surfaces produces cell proliferation and a marked increase in mitotic activity which may eventually lead to metaplasia and neoplasia.