Immunological Observations Research Articles

Heterotopic Heart–Unilateral Left Lung Transplantation in Dogs

A technique is presented for transplantation of the heart and left lung in the canine model. Preservation of the recipient's heart and right lung allowed heterotopic en bloc transplantation of the donor organs, thus preserving the normal respiratory pattern and obviating the need for cardiopulmonary bypass. Four experiments were performed to develop the surgical technique. The mean survival of the remaining 10 dogs studied was 23 ± 28 days (range 3 to 91 days). The transplanted heart and lung could be investigated as in orthotopic heart-lung transplantation; bronchoscopy with bronchoalveolar lavage and endobrachial biopsy as well as endomyocardial biopsy were performed by standard approaches. This model is suitable for physiological and immunological long-term observations of cardiopulmonary transplantation in dogs.

An Animal Model of Pulmonary Radiation Fibrosis with Biochemical, Physiologic, Immunologic, and Morphologic Observations

An animal model of pulmonary radiation fibrosis was established, using male CBA/j mice. Both lungs of each mouse in one group (DL) were irradiated with two doses of 8.5 Gy each, separated by 30 days. A control group (CG) was sham-irradiated. There was a small but significant difference (P less than 0.03) in average breathing rate between DL and CG 27 weeks after the second irradiation which increased until the 34th week followed by a plateau. The accumulated hydroxyproline content of the irradiated mouse lung was 40% greater (P less than 0.02) than that of the sham-irradiated lung at 42 weeks and thereafter. Anticollagen antibodies assayed 52 weeks after irradiation by enzyme-linked immunosorbent assay were elevated by 49% in sera from the irradiated mice compared to sera from sham-irradiated mice. Mortality during the 52-week period following the second irradiation was low (13%) for both groups. Histological comparison of irradiated and control mouse lungs fixed under uniform inflation pressure indicated no significant differences. The model has unique features including an increase in collagen deposition, no acute changes attributable to radiation, a small but statistically significant abnormality in pulmonary function, an immunologic response to collagen, and low mortality.

Immunological and clinical observations in diabetic kidney graft recipients pretreated with total-lymphoid irradiation.

In a feasibility study, twenty patients with end-stage diabetic nephropathy were treated with fractionated total-lymphoid irradiation (TLI, mean dose 25 Gy), before transplantation of a first cadaveric kidney. During radiotherapy, only one patient had a serious side effect (bone marrow depression). After transplantation four patients died (one of a myocardial infarction, one of ketoacidosis, and two of infections occurring during treatment of rejection crises). One graft was lost because of chronic rejection. The other 15 patients have a functioning graft (mean follow-up 24 months) and receive low-dose prednisone alone (less than 10 mg/day, n = 11) or in conjunction with cyclosporine (n = 4) as maintenance immunosuppressive therapy. A favorable clinical outcome after TLI (no, or only one, steroid-sensitive rejection crisis) was significantly correlated with a high pre-TLI helper/suppressor lymphocyte ratio, a short interval between TLI and the time of transplantation, and the occurrence of functional suppressor cells early after TLI. The most striking immunological changes provoked by TLI consisted of a long-term depression of the mixed lymphocyte reaction and of the phytohemagglutinin, and Concanavalin A or pokeweed-mitogen-induced blastogenesis. A rapid and complete recovery of the natural killer cell activity was observed after TLI. A permanent inversion of the OKT4+ (T helper/inducer) over OKT8+ (T suppressor/cytotoxic) lymphocyte ratio was provoked by a decrease of the OTK4+ subpopulation, together with a supranormal recovery of the OKT8+ lymphocytes. A majority of the latter lymphocytes did also express the Leu 7 and the Leu 15 phenotype.

HTLV-III antibody prevalence among young delinquent drug abusers in long-term residential treatment at a North-German drug clinic.

To date, long-term intravenous drug users studied at regional European centers of illicit drug use have had a high number of LAV/HTLV-III infections. Among 200 patients remanded by court or referred from prison to a special clinic in northern Germany for young delinquent drug abusers, 26 (17%) of 157 IV drug abusers were HTLV-III seropositive. With 40% seropositive, female patients showed a significantly higher prevalence of HTLV-III infection than males. The results of longitudinal serological, immunological, and clinical observations over periods of 12 months and 2-3 years indicate that under conditions of continuous medical surveillance, sound preventive counseling, and steady therapeutic care during long-term coeducative residential treatment of drug abusers, neither detectable HTLV-III transmission nor definite progression of HTLV-III induced impairment of immune regulatory functions must ensue.

Immunological and ultrastructural observations on swine thymic lymphoma

A case of swine thymic lymphoma is described. A young pig had a mediastinal mass and bone marrow involvement and the microscopic findings showed peripheral blood involvement. The tumour was composed of small to medium sized lymphoid cells which formed rosettes with sheep erythrocytes and were characterized by narrow cytoplasmic bands, nuclear convolution and clustered dense bodies. Such structural and immunological features resembled those of human thymic lymphomas derived from thymic T-cells.

Experimental infection of pregnant sheep with Toxoplasma gondii: Pathological and immunological observations on the placenta and foetus

The pathogenesis of Toxoplasma gondii infection in pregnant sheep was investigated. In one experiment, sheep were infected at 40, 60 and 90 days gestation and examined 30 days later, while in a second experiment, pregnant sheep were infected at either 60 or 90 days gestation and examined 10, 15 and 20 days later. By 10 days post infection (dpi), the parasite had invaded the gravid uterus and caused focal damage in the caruncular septa and, by 15 days, the degenerative changes had spread to the crypt wall and trophoblast. At this time, toxoplasms were isolated from all challenged foetuses. Foetuses did not produce any significant immune response until 20 dpi when increased numbers of IgM positive cells were found in lymphoid tissues. However, at 30 dpi, the numbers were normal while numbers of IgG positive cells and circulating IgG were increased. It was concluded that, while the foetal immune system need not be fully mature to respond to T. gondii infection, its ability to mount a response is limited at 70 days gestation but becomes increasingly competent after this. Furthermore, while it is able to respond more comprehensively with increasing maturity, maturation itself appears to be accelerated by the stimulus of infection.

Secondary immune response in the middle ear: immunological, morphological, and physiological observations.

A model of secondary immune response in the middle ear (ME) was developed in the guinea pig. Animals were immunized intradermally with keyhole limpet hemocyanin (KLH) and challenged with KLH in the ME 8 weeks later. The ME displayed effusion, mucosal hyperplasia, mucosal edema, and leukocytic infiltration. These responses were maximal at 3 days postchallenge, and persisted for 2 to 3 weeks. High levels of antibody were observed in the ME effusions, and serum titers increased significantly. Immunohistochemical localization of antibody indicated that substantial free IgG was present in the submucosa, with much less in the mucosal epithelium. IgA plasma cells were observed in the submucosa at 1 week postchallenge, but no IgM was detected. In contrast to the response observed in sensitized animals, antigen presented to the ME of unimmunized control animals resulted in no effusion, minimal mucosal inflammation, and little or no antibody in either the ME or serum. However, substantial neutrophilic infiltration of the ME cavity was observed. The effusion and inflammation produced by secondary ME immune responses were not affected by tympanostomy tubes. Eustachian tube function as measured by a forced response test was not affected by ME immune response. This suggests that the immunologically induced effusion was not the result of eustachian tube obstruction. We postulate that increased vascular permeability in the ME mucosa, induced by chemical mediators, resulted in the transudation of serum.

Clinical and immunologic observations in patients who stop venom immunotherapy

It is currently recommended that venom immunotherapy (VIT) be continued as long as the sensitivity persists (indicated by positive venom skin tests or RAST). In this pilot study, we performed a retrospective survey of the clinical and immunologic effects of stopping VIT. The 82 patients studied had received maintenance VIT for a mean of 14 months and had stopped VIT a mean of 43 months before evaluation. Subsequent “field” stings in 28 patients caused systemic reactions in six cases (22%), which is significantly higher than the 1% to 3% systemic reaction rate in patients who remain on maintenance VIT. The 22% reaction rate is a minimal estimate caused by loss of venom sensitivity in some patients and residual venom-specific IgG antibody levels in others. Reevaluation of venom skin tests and IgG levels was possible in 43 patients. A tenfold decline from before VIT skin test results was observed in 27 patients (63%). Skin tests remained clearly positive in 32/43 (74%), became weakly positive in 9/43 (21%), and 2/43 (5%) became negative. The IgG level declined from typical maintenance levels before stopping VIT (mean 7.2 ± 1.2 μg/ml) to levels typical of untreated patients at the time of retesting (mean 1.95 ± 0.3 μg/ml). Despite the marked fall of IgG antibody, one third of the patients still had levels in the average range observed in patients receiving maintenance VIT. We conclude that there is a substantial risk of anaphylactic sting reaction if VIT is stopped while venom sensitivity persists.

Strongyloides stercoralis: parasitological, immunological and pathological observations in immunosuppressed dogs

Four specific pathogen-free, two to three-month-old, laboratory-raised beagles were infected with Strongyloides stercoralis and monitored by thrice-weekly faecal examinations. Larvae were not detected in the faeces after the 11th week. During the 14th week, daily oral prednisolone (2·2 mg/kg) was given. This led to weak recrudescence of the infection. During the 21st week, the daily dose of prednisolone was doubled and thereafter two of the four dogs developed severe hyperinfective strongyloidiasis with their adult worm populations markedly exceeding the number of larvae inoculated, as did one dog which remained asymptomatic. One dog died during the 15th week, i.e., before there was time for a marked increase in the parasite population. Three of the four dogs provided evidence of disseminated infection with adult worms occurring in ectopic sites. Parasite-specific cellular and humoral immune responses were monitored weekly. All dogs developed a significant rise in specific IgM titres lasting until the fifth week. From the fourth week, all dogs had detectable parasite-specific IgG antibody levels that persisted throughout the experiment in spite of the immunosuppression. Strongyloides-induced lymphocytic responses in vitro were demonstrated in three dogs from the second to the fifth week of infection, but returned to pre-infection levels thereafter. All animals were necropsied. Relevant pathological lesions were found in the small intestine, the colon and the lungs. Larvae were seen in such ectopic sites as mesenteric nodes and prostate. The observations provide further evidence that canine S. stercoralis infections can be manipulated to reproduce with great fidelity the parasitological and clinico-pathological events occurring in human disseminated strongyloidiasis.

Morphological and Immunological Observations in Experimentally Induced Torsion of Testis in Rats

In an experimentally established model of torsion of the testis, morphological and immunological observations were evaluated in 180 Wistar rats. Torsion of the testis were carried out to observe in animals the biological phenomenon that naturally occurs in man. The effect of the twisted testis on the contralateral testis was carefully checked. The morphological observations revealed serious damage to the seminiferous tubules in the contralateral testis. In four rats, persistent infertility was observed as an effect of torsion. The infertility was confirmed by total atrophy of seminiferous epithelium in testis section. The cell-binding ability of sera obtained from rats at different times after torsion were studied by cell-binding radioimmunoassay with different types of germinal cells. The presence of autoantibodies to epididymal spermatozoa was revealed in most cases, being a secondary effect of the thus-far unknown, nonspecific factors responsible for damage of the contralateral testis.

Pathogenesis and immunology of chlamydial infections of the genital tract.

In recent years there has been a steady increase in our understanding of the pathogenesis and immunobiology of lower genital tract infections caused by Chlamydia trachomatis. Much of this increase in knowledge has stemmed from two areas of research: clinical, pathologic, and immunologic observations of patients and the development and study of animal models. Research on patients has included the use of techniques such as histologic examination of biopsy specimens and measurement of levels of antibody and/or leukocyte responses to chlamydial antigens. The animal models that have been developed include experimental genital tract infections not only with C. trachomatis but also with appropriate Chlamydia psittaci organisms, such as the agent of guinea pig inclusion conjunctivitis. Continued investigation of the pathophysiologic and immunologic bases of chlamydial genital tract disease should lead to the rational development of potential control measures, such as vaccines or other forms of prophylaxis or therapy.

Peripheral T Cell Lymphoma: Immunologic and Cell-Kinetic Observations Associated With Morphological Progression

Peripheral T cell lymphomas (PTCLs) form a morphologically heterogeneous group of non-Hodgkin's lymphomas that are generally considered to have immunophenotypes associated with mature T cells, usually those of helper T cells. We now describe and correlate the clinical, morphological, immunologic, and cell-kinetic findings based on the evaluation of eight tissue samples obtained at various times from a 13-year-old girl with PTCL. The early morphological expressions of this patient's PTCL were those of diffuse mixed-cell lymphoma and focal large-cell lymphoma (LCL) evolving from the histologic picture of an atypical immune response (AIR). These morphological findings were associated with an immature T cell immunophenotype associated with cortical thymocytes--namely, sheep erythrocyte rosette (sER)+, T11+, Leu-2a+, Leu-3a+, HLA-DR+, OKT6-, OKT9+, OKT10+--and with cell-kinetic findings that showed no evidence of aneuploidy and few cells in S phase. Diffuse pleomorphic LCL developed, which was associated with further dedifferentiation of the neoplastic T cells to the immunophenotype sER-, T11+, Leu-2a-, Leu-3a-, HLA-DR+, OKT6-, OKT9+, OKT10- and with cell-kinetic findings that demonstrated a distinct aneuploid population and a dramatic increase in the percentage of cells in the S phase. The immunophenotype of the PTCL at the time of the patient's death was T11-, Leu-2a-, Leu-3a-, HLA-DR+, OKT6-, OKT9+, OKT10-, an immunophenotype indistinguishable from that of a non-B non-T cell lymphoma. The immunologic findings in this case also suggest that an AIR in some cases may represent a prelymphomatous state or may be a morphological expression of PTCL. These observations indicate that PTCLs may be characterized by rapidly changing clinical, morphological, immunologic, and cell kinetic findings which are best evaluated by multidisciplinary studies.

Peripheral T cell lymphoma: immunologic and cell-kinetic observations associated with morphological progression

Peripheral T cell lymphomas (PTCLs) form a morphologically heterogeneous group of non-Hodgkin's lymphomas that are generally considered to have immunophenotypes associated with mature T cells, usually those of helper T cells. We now describe and correlate the clinical, morphological, immunologic, and cell-kinetic findings based on the evaluation of eight tissue samples obtained at various times from a 13-year-old girl with PTCL. The early morphological expressions of this patient's PTCL were those of diffuse mixed-cell lymphoma and focal large-cell lymphoma (LCL) evolving from the histologic picture of an atypical immune response (AIR). These morphological findings were associated with an immature T cell immunophenotype associated with cortical thymocytes--namely, sheep erythrocyte rosette (sER)+, T11+, Leu-2a+, Leu-3a+, HLA-DR+, OKT6-, OKT9+, OKT10+--and with cell-kinetic findings that showed no evidence of aneuploidy and few cells in S phase. Diffuse pleomorphic LCL developed, which was associated with further dedifferentiation of the neoplastic T cells to the immunophenotype sER-, T11+, Leu-2a-, Leu-3a-, HLA-DR+, OKT6-, OKT9+, OKT10- and with cell-kinetic findings that demonstrated a distinct aneuploid population and a dramatic increase in the percentage of cells in the S phase. The immunophenotype of the PTCL at the time of the patient's death was T11-, Leu-2a-, Leu-3a-, HLA-DR+, OKT6-, OKT9+, OKT10-, an immunophenotype indistinguishable from that of a non-B non-T cell lymphoma. The immunologic findings in this case also suggest that an AIR in some cases may represent a prelymphomatous state or may be a morphological expression of PTCL. These observations indicate that PTCLs may be characterized by rapidly changing clinical, morphological, immunologic, and cell kinetic findings which are best evaluated by multidisciplinary studies.

Absence of Antibodies to HTLV-III in Health Workers After Hepatitis B Vaccination

A proportion of the plasma for the triply inactivated, plasma-derived hepatitis B vaccine produced in the United States is obtained from homosexual men. Because homosexual men are a high-risk group for the acquired immunodeficiency syndrome (AIDS), concern has emerged that the vaccine could harbor the AIDS agent. To evaluate this risk, we tested 15-month postvaccination serum samples for antibodies to human T-cell lymphotropic virus type III in 100 health care workers who had received inactivated hepatitis B vaccine lots made from plasma collected between 1977 and 1979 and 100 who had received placebo injections. None of the 200 health workers had serological evidence of human T-cell lymphotropic virus type III infection. These serological findings lend additional support to earlier epidemiologic and immunologic observations suggesting that hepatitis B vaccine does not transmit infection with an AIDS virus.

AIDS and the gut.

The epidemiological, immunological and early virological observations on the acquired immune deficiency syndrome (AIDS) suggested that an agent was involved which was sexually, parenterally and perinatally transmitted and perhaps tropic for T helper lymphocytes. A new subgroup of human T lymphotropic retroviruses have been identified ans seroepidemiological studies suggest that they are aetiologically related to AIDS. The syndrome is characterised by the development of tumors: such as Kaposi's sarcoma and non-Hodgkins lymphoma, with an aggressive clinical course and infection by a wide spectrum of opportunistic organisms. Both the tumours and the infections commonly involve the gut.

Immunologic observations in sera of a patient with allergic bronchopulmonary aspergillosis by means of the enzyme-linked immunosorbent assay

Titers of IgG, IgA, and IgM antibodies against Aspergillus fumigatus were measured by ELISA in sera of a patient with allergic bronchopulmonary aspergillosis during an acute stage, remission, and exacerbation of the disease. Total IgE, specific IgE against A. fumigatus, and number of precipitation lines were also measured. ELISA IgG-, IgA-, and IgM-antibody titers were expressed in percentages of the titer of a strongly positive standard serum by use of the patient's own serum as a standard. The fall in antibody concentration (in percent) and elevations during exacerbation appeared to be similar for these antibody classes and are related with the number of precipitates measured by double immunodiffusion. Furthermore, the changes in antibody titers were also similar to the elevations and decreases in both total IgE and specific-IgE antibodies (also expressed in percentages of a chosen standard serum). The ELISA IgG-antibody titers were higher compared with the ELISA IgA or IgM titers and demonstrated a continuous decline during a period of 2 yr after the acute phase of ABPA. Therefore, IgG-antibody titers are preferred as an indicator of disease activity in follow-up studies over longer time periods and are at least as sensitive as the measurement of concentrations of IgE antibodies.

Pleural Mesothelioma and Neighborhood Asbestos Exposure

IN 1978, we reported a case of malignant pleural mesothelioma with unusually long survival. In that report we detailed histopathologic and immunologic observations that may have been related to the long-term survival.¹In this article, we summarize the clinical course and relate the probable etiology of the disease to an unusual type of asbestos exposure. <h3>Report of a Case</h3> A male patient, born in 1924, was well until the beginning of 1970, when he noted a gradual increase in shortness of breath and sharp pain in the right side of the chest. He experienced repeated pleural effusions and was found to have malignant pleural mesothelioma. He was not given chemotherapy or any specific treatment for the disease, which showed a very slowly progressing course. In December 1979, there was a rapid deterioration in the patient's clinical status, with marked progress of the chest roentgenographic abnormalities (Fig 1). The patient

Human placental beta-galactosidase: structural and immunological observations.

beta-Galactosidase purified to apparent homogeneity from human placenta occurred in two separable fractions. A low molecular mass form (relative mass (Mr) 170 000) is composed of a single polypeptide chain (Mr 70 000). This was derived from a larger form by molecular sieve chromatography at both low (10 mM) and high (500 mM) NaCl concentration. The larger form of beta-galactosidase also contains small amounts of two polypeptides with apparent Mr values of 23 000 and 35 000 daltons. Both forms of the enzyme hydrolyze synthetic aryl galactosides and natural glycolipid substrates at comparable rates. Antibodies raised in rabbits against the low Mr beta-galactosidase also cross-reacts with the high Mr enzyme. The antibody preparation also cross-reacted with beta-hexosaminidase even though the latter was found at very low levels in the antigen, as judged by lack of detection of representative protein bands on sodium dodecyl sulfate - polyacrylamide gel electrophoresis and enzyme activity measurements. A portion of this cross-reactivity (35%) against beta-hexosaminidase could not be absorbed from the preparation without the simultaneous loss of beta-galactosidase activity, suggesting that the two enzymes show a degree of antigenic identity.

Virologic, immunologic, and clinical observations on a patient during the incubation, acute, and convalescent phases of infectious mononucleosis

One patient with infectious mononucleosis (IM) was studied from the probable time of Epstein-Barr virus (EBV) infection (38 days before the onset of clinically overt disease), during the incubation and acute phases, until 6 months after clinical remission. Analysis of spontaneous outgrowth of EBV-carrying lymphoblastoid cells, by limiting dilution on feeder layer cultures, showed that virus containing B lymphocytes are already present early during the incubation period. Also low interferon serum levels were detected early after infection, and only before the onset of clinical disease. All other studied clinical laboratory and virus-associated variables were within normal range during the incubation phase, but changed to a pattern characteristic of IM in parallel to the clinical symptoms. During the acute disease EBV-associated nuclear antigen (EBNA)-positive cells could be directly detected among the lymphocytes, and antibodies to EBV antigens appeared. Lymphocytes stained by monoclonal antibodies, detecting Ia-like determinants (activated cells) and supressor cells, increased dramatically, in parallel to a strong increase of functional suppressor cell activity, measured by inhibition of blastogenesis and PWM-induced immunoglobulin production. During the acute phase there was also a decrease of spontaneous cytotoxicity against the NK-sensitive cell line K562, while cytotoxicity (spontaneous) against an autologous EBV-positive lymphoblastoid cell line (LCL) was detected only during this phase. These reactions correlated to the presence of blasts, and the autologous reaction was exerted mainly by Fc-receptor-negative cells. Lymphokine production in response to EBV antigens was also initiated during the acute phase. During the convalescence period the serological and cellular immune parameters adjusted to the pattern of a normal EBV-seropositive person.

Molecular analysis of immunoglobulins M and G immune response to protein antigens of Treponema pallidum in human syphilis.

Protein antigens of Treponema pallidum precipitated by immunoglobulin M (IgM) and IgG antibodies of sera from patients with untreated primary and secondary syphilis as well as treated secondary syphilis were characterized on a molecular basis. T. pallidum was labeled internally with [35S]methionine and solubilized in 0.1% sodium dodecyl sulfate-1% Triton X-100. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 12.5% gels followed by autoradiography revealed 32 distinct proteins with molecular weights between 13,500 and 200,000. Twenty-three proteins of T. pallidum with molecular weights between 15,500 and 115,000 were identified as antigens by double antibody radioimmunoprecipitation with IgM and IgG antibodies of sera from syphilitic patients. The molecular analysis of the IgM and IgG immune response to T. pallidum in human syphilis is in accord with earlier immunological observations. Finally, utilizing syphilitic human sera, we characterized 15 protein antigens of T. pallidum that are common to Treponema phagedenis by partial absorption of IgM and IgG antibodies with an ultrasonicate of T. phagedenis.