Kunal Saha

Immunological studies in thromboangiitis obliterans (Buerger's disease)

Abstract Immunological studies were carried out in 10 patients of thromboangiitis obliterans (Buergers disease) and compared with controls. Cell mediated and humoral immunities were investigated. Presence of specific cellular immunity against arterial antigen, increased serum immunoglobulins, formation of specific humoral antiarterial antibodies, and demonstration of immune complexes raise the possibility of involvement of autoimmune process in causation of thromboangiitis obliterans.

Autoantibodies in thromboangiitis obliterans (Buerger's disease).

Autoantibodies, organ and non-organ specific, were studied in 10 patients of thromboangiitis obliterans by immunofluorescence technique and compared with age matched controls without peripheral vascular disease and patients of other peripheral vascular diseases. Organ specific autoantibodies (various classes of immunoglobulins IgM, IgG, IgA) and C3 component were observed in the diseased vessels of thromboangiitis obliterans. Antibodies, antiarterial, were present in the sera of these patients. Among the non-organ specific antibodies (ANA and AMA), only ANA were found in the sera of patients. Presence of autoantibodies in patients of thromboangiitis obliterans strongly point out of that probably this disease entity is an autoimmune disorder.

BIOLOGICAL VARIATION OF IMMUNOGLOBULIN CONCENTRATIONS IN NORMAL HUMAN TEARS RELATED TO AGE AND SEX

Single radial immunodiffusion method was used to measure the concentration of IgG, IgA, IgM and IgD in tears of 220 healthy individuals aged from 2 to 86 years. Relation of the values to age and sex has been evaluated statistically by regression analysis method. Mean IgA level was 30.7 mg/100 ml. IgG could be detected in 200 samples and the level was less than 1 mg/100 ml. IgM was detected in only 7 samples and the value was less than 1 mg/100 ml. IgD could not be detected in any of the sample. The IgA level in males and that in females differs significantly, the females having a higher mean value. The IgA level appears to increase in both sexes with age. No relationship with age and sex could be established in other types of immunoglobulins.

Treatment of Patients with Thromboangiitis Obliterans with Cyclophosphamide

This report describes a small, nonrandomized trial of cyclophosphamide in the treatment of patients with advanced thromboangiitis obliterans (TAO) with modest results. The rationale of the treatment was based on the immunopathogenesis of the disease, ie, autoimmune vasculitis of peripheral arteries. Twelve male patient volunteers with TAO were included for the trial. Diagnosis was based on the history of chronic smoking or tobacco chewing, clinical features of ischemia of peripheral vessels, radioarteriography showing arterial block, and characteristic histopathologic changes of affected arteries. Cyclophosphamide (400 mg) was given intra venously daily to the patients for 7 days followed by daily oral administration of 100 mg cyclophosphamide for another 7 weeks. Clinical conditions of the patients started to improve during the third week of the treatment and maximum benefit was noticed at the end of the treatment. There was significant decrease of intermittent claudication and twentyfold increase of claudication distance as well as relief of rest pain. Before starting treatment 6 patients had developed ulcers on their affected limbs; these healed completely in 2, partially healed in another 2, and showed no improvement in the remaining 2, who never stopped smoking. However, immunosuppressive therapy failed to show any improvement of arterial block, as evidenced by radioarteriography and any significant increase of skin temperature over the affected limbs. Nevertheless, histopathologic studies of biopsies taken from the diseased arteries after completion of therapy showed decreased influx of lymphocytes and plasma cells in the thrombi as well as in the arterial walls in comparison to the biopsies taken before the start of treatment. During the treatment the degree of immunosuppression was monitored by blood leukocyte and lymphocyte counts, which were kept between 4,000/mm3 and above 3,000/mm3 and not less than 500/mm 3, respectively, indicating modest immunosuppression and no serious complications. All patients were followed up for 1 year. Only 2 patients, who resumed smoking, had relapse.

Significance of circulatory immune complexes in thromboangiitis obliterans (Buerger's disease).

Circulatory immune complexes were estimated in the sera of 25 patients and 20 age and sex matched controls by precipitation with 8% polyethylene glycol (PEG). Characterization of the immune complexes, thus obtained, was performed by the double immuno-diffusion method. The mean protein content of the PEG precipitates was significantly higher in the patients of thromboangiitis obliterans than in the controls. The immunoglobulins in the immune complexes were predominantly of mixed type in contrast to mainly single class immunoglobulins in the controls. C3 complement distribution in immune complexes of patients and control sera was 64% and 45% respectively. The immune complexes in patients were found to be biologically active. The mechanism of formation of circulatory immune complexes and their role in the pathogenesis of arteritis in relation to smoking of tobacco has been postulated.

A study of serum immunoglobulins including IgE and reaginic antibodies in patients with bronchial asthma in relation to the spectrum of the disease

Immunoglobulins G, A, M, D, and E were studied in the sera obtained from sixty‐nine bronchial asthma patients, who were graded objectively according to atopic score. Among other associated atopic diseases, they often had allergic rhinitis. Ninety‐one per cent of these patients had elevated serum IgE levels and the mean level of serum IgE was more than 3.5 times higher than that observed in the normal subjects. Moreover, as their atopic score increased, the serum IgE levels were also elevated, and every patient with high atopic score, had elevated serum IgE levels: This indicates the association of atopic state with serum IgE level.

Natural resistance against tetanus in patients with lepromatous leprosy

Tetanus antitoxin levels in the sera of unimmunized lepromatous patients, often suffering from many ulcerations on their limbs due to physical and thermal trauma, but not developing clinical tetanus, were studied. Sera from 40 such patients with no history of immunization against tetanus, 35 unimmunized matched controls and 12 immunized controls were tested for the presence of tetanus antitoxin by the passive haemagglutination technique. 45% of patients, 17% of controls and all the immunized patients showed adequate serum antibody levels. Eight of the 22 leprosy patients, with no detectable antitoxin on their sera, were immunized with a single dose of tetanus toxoid and all showed sero-conversion four weeks later. 2-Mercaptoethanol (2ME) treatment of these sera showed both IgM and IgG classes of specific antibody in all groups but the level of mature antibody of IgC class was higher in the immunized group. Neutralization test further confirmed the presence of protective antibodies in these sera.

High incidence of IgG class of Epstein-Barr virus capsid antibody in Indian patients of lepromatous leprosy

Low levels of Epstein-Barr virus capsid (EBVC) antibody of the IgG class were detected in the sera of 19 of 23 (82.6%) lepromatous leprosy patients and six of 38 (16%) healthy controls. In contrast, heterophile antibody was found in only six of 43 (14%) lepromatous patients and three of 41 (7%) normal subjects. Overlap of the two types of antibody occurred only in one normal serum. It is inferred that the presence of EBVC antibody against an unbiquitous virus in lepromatous patients who often suffer from impairment of cell-mediated immunity might be due to past infection leading to persistence of the virus in their lymphoid cells and subsequent production of specific anti-viral antibody. Further, the striking finding of low incidence and titre of EBVC antibody in the normal Indian adults is consistent with the rarity of EB virus-associated disease, such as Burkitts lymphoma, nasopharyngeal carcinoma and infectious mononucleosis in India.

Post-transfusion hepatitis and hepatitis-associated antigen in New Delhi

Abstract A study of 58 random recipients of single or multiple blood transfusions revealed post-transfusion hepatitis in 18·9%, based upon clinical and biochemical assessment. Using counter-immunoelectrophoresis, hepatitis associated antigen (HAA) alone was detected in 27·2% of cases, HAA and/or anti-HAA was found in 45·4% cases of hepatitis, and 13·7% of all the recipients. Out of 8 patients with HAA and/or anti-HAA, 3 remained normal clinically as well as biochemically, while the remaining 5 developed either icteric or anicteric hepatitis. Of the latter, one subject was associated with both antigen and antibody, 2 with antigen and2 with antigen and 2 with antibody alone. Among 306 asymptomatic blood donors, HAA and anti-HAA were found in 16 (5·2%) and 2 (0·65%) cases respectively, without any striking difference in the percentage of HAA in voluntary and professional donors. Of the 102 donors tested for transaminases, 4 showed a minimal abnormality in transaminases, only 2 of which were associated with the antigen. A control group of 1,156 subjects showed antigen in 2·76%.

STATUS OF FIBRIN DEGRADATION PRODUCTS IN LEPROSY

Plasmafibrinogen and fibrin degradation products (FDP) were estimated in 45 control subjects and 45 patients with various types of leprosy including 16 patients with erythema nodosum leprosum (ENL). The levels of FDP were quantified by using a suspension of Staphylococcus aureus (Newmans D2C strain). Significantly higher levels of plasma fibrinogen and FDP were observed in patients with lepromatous leprosy, especially with ENL, than in the controls. Only 3 lepromatous patients had plasma fibrinogen levels below the normal range; 2 of them had severe ENL and were suffering from nasal bleeding. Levels of D and E fragments were also estimated in another 20 control subjects and 13 lepromatous patients by haemagglutination inhibition technique and were found to be significantly elevated in the patients. It was inferred that raised FDP levels in such patients indicated an ongoing occult fibrinolysis during lepra reaction. A few lepromatous patients may show hypofibrinogenomia during severe ENL episodes.