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Biomedical subjects

T B Nutman

Publications and source records attributed to T B Nutman.

At least 163 records · Page 9Linked to original sources
PubMed1987

Parasite-specific anergy in human filariasis. Insights after analysis of parasite antigen-driven lymphokine production.

The antigen-specific immune unresponsiveness seen in bancroftian filariasis was studied by examining lymphokine production in peripheral blood mononuclear cells (PBMC) or PBMC subpopulations from 10 patients with asymptomatic microfilaremia, 13 patients with elephantiasis and 6 normal North Americans. In each group of patients, the kinetics of the lymphokine response and the response to mitogens and nonparasite antigens did not differ significantly. In marked contrast, when antigen-induced lymphokine production was examined, most patients with microfilaremia were unable to produce either interleukin 2 (IL-2) or gamma-interferon (i.e., were nonresponders), and the few who could (hyporesponders, generally with quite low microfilaremia levels) did so at levels significantly less than those of patients with elephantiasis, all of whom showed strong responses to parasite antigen. Removal of neither adherent cells or T8+ cells affected the parasite-specific anergy seen in those with microfilaremia, suggesting a state of T cell tolerance to the parasite in patients with this most common clinical manifestation of bancroftian filariasis.

Adult↗
PubMed1987

Acute tropical pulmonary eosinophilia. Characterization of the lower respiratory tract inflammation and its response to therapy.

Although acute tropical pulmonary eosinophilia (TPE) is well recognized as a manifestation of filarial infection, the processes that mediate the abnormalities of the lung in TPE are unknown. To evaluate the hypothesis that the derangements of the lower respiratory tract in this disorder are mediated by inflammatory cells in the local milieu, we utilized bronchoalveolar lavage to evaluate affected individuals before and after therapy. Inflammatory cells recovered from the lower respiratory tract of individuals with acute, untreated TPE (n = 8) revealed a striking eosinophilic alveolitis, with marked elevations in both the proportion of eosinophils (TPE 54 +/- 5%; normal 2 +/- 5%; P less than 0.001) and the concentration of eosinophils in the recovered epithelial lining fluid (ELF) (TPE 63 +/- 20 X 10(3)/microliter; normal 0.3 +/- 0.1 X 10(3)/microliter; P less than 0.01). Importantly, when individuals (n = 5) with acute TPE were treated with diethylcarbamazine (DEC), there was a marked decrease of the lung eosinophils and concomitant increase in lung function. These observations are consistent with the concept that at least some of the abnormalities found in the lung in acute TPE are mediated by an eosinophil-dominated inflammatory process in the lower respiratory tract.

Adult↗
PubMed1987

Antibodies to porcine eye muscle in patients with Graves' ophthalmopathy: identification of serum immunoglobulins directed against unique determinants by immunoblotting and enzyme-linked immunosorbent assay.

The immunological mechanisms involved in Graves' ophthalmopathy are not known. To explore the pathophysiology of this disorder, we used an enzyme-linked immunosorbent assay to detect antibodies that bound to the 100,000 X g sediment fraction of porcine eye muscle. Serum from patients with Graves' ophthalmopathy had enhanced immunoglobulin binding to porcine eye muscle compared to serum from control subjects (P less than 0.0001); however, several individual normal serum samples also had elevated binding activity. Incubation of serum from some Graves' patients with skeletal muscle or liver tissue resulted in reduction in immunoglobulin binding to porcine eye muscle. Thyroglobulin and TSH reduced binding only at high concentrations. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed some obvious differences in protein bands between porcine eye muscle and skeletal muscle. Serum from many normal subjects and patients with autoimmune thyroid disease formed bands with skeletal muscle protein fractions on immunoblotting, and no specific bands were found using serum from Graves' ophthalmopathy patients. Immunoblots of Graves' patients' serum after reaction with either porcine skeletal or eye muscle showed no reactivity with thyroglobulin (200K and 320K) or thyroid microsomal antigen (105K). Instead, immunoblots of eye muscle and serum from Graves' patients, with or without eye disease, showed two bands at 64K and 73K, which were shared by serum from normal subjects. In addition, several unique eye muscle determinants were detected in serum from 6 of 13 Graves' disease patients analyzed. These unique bands usually had mol wt of less than 50K, and 5 of the 6 patients whose serum reacted with these determinants had significant ophthalmopathy. Our findings support the presence of potential antigenic differences between eye muscle and skeletal muscle accounting for the immunological specificity of eye muscle as a target in Graves' ophthalmopathy.

Animals↗
PubMed1987

The clinical and immunologic responses of normal human volunteers to low dose hookworm (Necator americanus) infection.

Five normal human volunteers were exposed to approximately 50 infective larvae of Necator americanus and were observed for the development of clinical signs or symptoms and for changes in blood eosinophil levels, IgG antibody titers, total and parasite-specific IgE, and lymphocyte blastogenic responses for 6-10 weeks. Bronchoalveolar lavage was performed on four subjects prior to infection and at times when larval migration through the pulmonary tree was likely. Eggs were demonstrated in the stools of four volunteers who remained untreated for more than 6 weeks; one volunteer had to be treated at day 40 because of severe gastrointestinal symptoms. All others also complained of abdominal pain and flatulence between days 35-40. All volunteers developed marked blood eosinophilia which peaked between days 38-64 and ranged from 1,350-3,828 eosinophils/mm3. Small increases in total and parasite-specific IgE and IgG were noted in some volunteers. One volunteer showed a significant lymphocyte blastogenic response. With the exception of mucosal erythema, bronchoalveolar lavage results were unremarkable. Our data indicate that a single small inoculum of hookworm larvae is capable of producing significant transient gastrointestinal morbidity and marked blood eosinophilia but does not induce other prominent T cell- and B cell-dependent immune responses.

Adult↗
PubMed1987

Persistent thyroid autoimmunity after subacute thyroiditis.

Subacute (de Quervain's) thyroiditis (SAT) is accompanied by temporary abnormalities in thyroid function and, in a minority of patients, by transient thyroglobulin and thyroid microsome autoantibody formation. In this report we have examined the sera of patients with SAT for the presence of multiple thyroid autoantibodies using qualitative immunoblotting (Western blotting). For this purpose we used a crude thyroid extract (2,000 g supernatant) as a source of antigen, in order to identify antibodies reacting with a wide range of potential autoantibodies. Eight of the 9 patients tested had autoantibodies which reacted with between 2-18 antigenic determinants (molecular weights 177-10 kd) present in the thyroid antigen preparation. None of the sera reacted with thyroglobulin or thyroid microsomes. The majority of these autoantibodies were directed against antigens which were absent from liver membrane preparations. Furthermore there was no diminution in the levels of these autoantibodies over a period of up to 39 months after the onset of SAT. The prolonged presence of these novel thyroid autoantibodies may explain the recent findings of subtle thyroid defects, such as altered gland iodine content, long after apparent clinical resolution of SAT. The accumulated evidence for a viral aetiology in this condition suggests that these sequelae may be due to a virally-induced autoimmune response.

Adult↗
PubMed1986

Loa loa infection in temporary residents of endemic regions: recognition of a hyperresponsive syndrome with characteristic clinical manifestations.

Loiasis in residents of endemic areas is generally manifested by microfilaremia, episodic angioedema, periocular migration of adult worms, modest eosinophilia, and variable antibody levels. In 20 temporary residents who acquired loiasis in West Africa, however, the clinical presentations were markedly different. Only three of the 20 had detectable microfilaremia. Furthermore, these patients often showed a state of marked immunological hyperresponsiveness manifested by very high titers of antibody to filariae, increased levels of serum IgE, and profound hypereosinophilia--to levels greater than 3,000/mm3 in 18 of the 20. These patients were also notable for the increased severity, frequency, and pruritic nature of their angioedema. Also, significant complications occurred in seven of the 20; one patient developed endomyocardial fibrosis and six, renal disease after treatment with diethylcarbamazine. Thirteen of the 20 developed subcutaneous nodules; adult parasites were recovered from three of five of these patients who underwent biopsy. All of the patients responded to therapy with diminution of their clinical symptoms and decreases in levels of eosinophils, IgE, and antibody to filariae.

Adult↗
PubMed1986

Identification of circulating parasite antigen in patients with bancroftian filariasis.

Because many cases of lymphatic filariasis cannot be diagnosed either clinically or by immunodiagnostic test based on antibody detection, recent efforts have been more directed towards developing methods for detecting parasite antigen in the blood or urine. Using a solid phase (Sepharose 4B) two-site immunoradiometric assay (IRMA) employing hyperimmune rabbit antifilarial antisera, we have previously shown (Hamilton et al., 1984) that essentially all cases of patent (ie. microfilaremic) infection in patients with bancroftian filariasis can be detected by this semi-quantitative assay as well as some individuals with amicrofilaremic (i.e., 'cryptic') infection. The present communication reports the results of studies that identify a prominent circulating antigen detected by this IRMA in sera from patients with microfilaremia. The antigen was eluted from Sepharose-bound rabbit polyclonal antiserum that had been reacted with known antigen positive sera. It was run in SDS-PAGE, blotted to nitrocellulose paper and identified autoradiographically using 125I-labelled rabbit antifilarial antiserum. Its high molecular weight (approximately 200 kD), stability to acid and boiling, and sensitivity to pronase and periodate suggest its being a glycoprotein. Isolation of this antigen will permit the development of specific reagents (such as monoclonal antibodies) which should enhance both the sensitivity and utility of the currently available antigen detection systems.

Adolescent↗
PubMed1985

In vitro parasite antigen-induced antibody responses in human helminth infections.

Helminth parasites characteristically induce vigorous antibody responses in human infections, but the immunoregulatory mechanisms determining the level of these responses are not at all understood. To investigate these mechanisms, peripheral blood mononuclear cells were obtained from 10 patients with parasitic helminth infections (three with schistosomiasis, three with onchocerciasis, and four with loiasis), along with six normal controls. These cells were then cultured in vitro and the capacity of the cells to respond to a specific parasite antigenic stimulus was examined by measuring the amount of parasite-specific antibody produced. Parasite antigen alone, without exogenous mitogen, induced an IgG anti-parasite antibody response in vitro. Optimal responses were obtained at extremely low antigen concentrations--concentrations at which little if any polyclonal immunoglobulin production occurred. Additionally, the in vitro induction of parasite-specific antibody was antigen dose-dependent, requiring much lower antigen concentrations than those necessary to induce lymphocyte blastogenesis. Antibody production was shown to require the cooperative interaction of B and T cells. These studies demonstrate that in vitro responses to antigens from naturally acquired parasitic infections, like those in individuals postimmunization, can be utilized to dissect the cellular and humoral factors that regulate antibody production to naturally acquired human pathogens.

Adult↗
PubMed1985

Filarial parasite-specific T cell lines: induction of IgE synthesis.

The development of T lymphocyte lines and clones of defined specificity has provided an important method for investigating T cell recognition of foreign antigens as well as T cell influence on B cell activity. We described previously a parasite-specific T cell line (TCL) derived from a patient with a naturally acquired filarial infection and elevated levels of serum IgE. The TCL is composed of Leu-3+ helper cells and is maintained independent of exogenous growth factors. In the present study, we used these T cells to investigate their immunoregulatory function on the in vitro IgE response. These parasite-specific T cells can provide isotype-specific help for antigen-induced IgE production by B cells in vitro. Autologous T cells profoundly suppress IgE production in a concentration-dependent manner. Furthermore, soluble factors generated from these filarial-specific TCL after antigen stimulation are able to induce the production of IgE in normal human cells not already synthesizing measurable amounts of IgE in vitro. Partial physicochemical characterization of this factor has shown that it is heat labile, has an m.w. between 10,000 and 30,000 M(r), and is a mannose-rich glycoprotein.

Absorption↗
PubMed1985

Production of monoclonal cyclic GMP and cyclic AMP antisera.

In order to facilitate the in situ localization of cyclic nucleotides, a large supply of high-titer anti-sera for immunohistologic analyses would be valuable. Therefore, monoclonal antibodies (Mab) directed at 2'-0-succinyl cyclic GMP (s-cGMP) or 2'-0-succinyl cyclic AMP (s-cAMP) were generated. Immunized mice developed polyclonal antibodies of high titer to either cyclic nucleotide as reflected in an enzyme-linked immunosorbent assay (ELISA) employed to monitor these anti-sera. With the use of standard hybridoma technology and ELISA to monitor anti-cyclic nucleotide antibody production from positive clones, several Mab-producing clones were grown to large volume in tissue culture and ascites fluid. The anti-cGMP and anti-cAMP anti-sera, which demonstrated significant ELISA titers at greater than 1:2,000,000 dilutions, are monospecific and can be employed for radioimmunoassays, ELISA, or immunohistologic localization of cyclic nucleotides in situ.

Animals↗
PubMed1984

Parasite antigen-specific human T cell lines and clones. Major histocompatibility complex restriction and B cell helper function.

The development of T lymphocyte lines and clones of defined specificity has become an important method for investigating both T cell recognition of foreign antigens as well as T cell influence on B cells. In the present study, human antigen-specific T cell lines and clones have been derived from a patient with a naturally acquired filarial infection. These T cells are of the helper phenotype (Leu 1+, Leu 2-, Leu 3+) and are independent of exogenous interleukin-2. Furthermore, these T cells have been shown to require both antigen-presenting cells and antigen for optimal proliferation. Helper function mediated by these T cells as manifested by the in vitro induction of parasite-specific antibody was antigen-dose dependent, requiring much lower antigen concentrations than those necessary to induce blastogenesis. More importantly, there is an absolute requirement of the T cell line for HLA-DR histocompatible antigen-presenting cells; clones derived from this T cell line show a more specific DR-related restriction--to only one of the two parental DR haplotypes in antigen stimulated proliferative responses. Such parasite antigen specific human helper T cell lines and clones should prove useful in exploring the fine control of the host response to naturally acquired helminth infections. In addition, these long-term T cell lines and clones can provide a potent tool for examining not only the events involved in human T cell responses to parasite antigens, but also into the associated cellular and humoral factors necessary for the B cell responses which follow.

B-Lymphocytes↗
PubMed1984

Differentiation stimuli induce receptors for plasma fibronectin on the human myelomonocytic cell line HL-60.

Plasma fibronectin (Fn) induces phagocytosis of C3b-opsonized sheep erythrocytes (EC3b) by human peripheral blood monocytes. However, Fn does not induce erythrophagocytosis of EC3b by human polymorphonuclear leukocytes (PMN), unless the PMN have been exposed to C5a or N-formyl-methionyl-leucyl-phenylalanine. Because of this difference, it is of great interest to examine Fn binding to cells that possess the capacity to differentiate into either granulocytes or monocytes. Hence, we have examined the consequences of Fn binding to the human myelomonocytic cell line, HL-60, both before and after in vitro differentiation of the HL-60, along a monocytoid or a granulocytoid pathway. Fn receptors were not found on undifferentiated HL-60, but several differentiating agents promoted the HL-60 binding of Fn-coated microspheres (Fn-ms). The peak of Fn-ms binding occurred four to five days after the induction of differentiation with dimethylsulfoxide (DMSO), and two days after induction by PMA. In addition, cells that differentiated along either the monocytoid or the granulocytoid pathway showed a marked increase in the phagocytosis of both IgG-coated erythrocytes (EA) and EC3b when they were exposed to Fn. Comparison of the effects of anti-Fn monoclonals on the binding of Fn-ms to the monocytes, PMN, and HL-60 showed that the same monoclonals block Fn-ms-binding and Fn-induced EC3b phagocytosis by all three cell types. Two monoclonal antibodies, M1/70 and A6F10, directed against membrane antigens on PMN and monocytes, inhibited Fn-ms binding. Both also blocked Fn-induced EC3b ingestion by these cells. However, neither antibody blocked Fn-ms binding or EC3b ingestion by differentiated HL-60. We conclude that differentiated HL-60 cells express functionally active Fn receptors, similar to monocytes and activated PMN, which, nonetheless, differ from normal cells in their association with the antigens recognized by M1/70 and A6F10.

Animals↗
PubMed1984

IgE production in vitro by peripheral blood mononuclear cells of patients with parasitic helminth infections.

Helminth parasites induce production of high levels of IgE antibodies but the immunoregulatory mechanisms determining this IgE biosynthesis are poorly understood. To investigate these mechanisms, peripheral blood mononuclear cells were obtained from six normal controls, six atopic patients and eight patients with parasitic helminth infections (three with schistosomiasis, two with loiasis, three with onchocerciasis). Cells were cultured at 1 X 10(6) cells/ml for 8 days in the presence of media alone or media supplemented with pokeweed mitogen (PWM) or cycloheximide; the supernatant fluids from these cultures were then assayed quantitatively for total and parasite specific IgE and IgG using an avidin-biotin amplified (for IgE) or standard (for IgG) microelisa assay. The geometric mean spontaneous IgE production was markedly elevated in peripheral blood mononuclear cells from parasitized individuals (2,487 pg/ml) when compared to those from atopics (358 pg/ml) or normals (152 pg/ml). Spontaneous IgG synthesis was equivalent in all three groups (range 140-420 ng/ml). PWM did not induce IgE production in any group and in the parasitized group even caused significant suppression of total IgE synthesis. Antigen specific antibody production (both IgE and IgG) paralleled total immunoglobulin synthesis. These findings demonstrate for the first time spontaneously enhanced IgE production in vitro in patients with helminth infections and provide a model system for studying the suppressive and regulatory mechanisms controlling IgE secretion.

Adolescent↗
PubMed1981

Lymphokine-mediated suppression of chondrocyte glycosaminoglycan and protein synthesis.

Spontaneously released and T cell mitogen augmented lymphokine produced by human mononuclear cells has been shown to induce a concentration dependent reversible suppression of chondrocyte glycosaminoglycan and protein synthesis without significantly enhancing chondrocyte catabolic activity. The modulatory factor(s) is of T cell origin and is trypsin, pronase, and heat sensitive. Prostaglandin inhibitors failed to influence factor formation or activity. Although eluting from Sephadex G-100 over a wide range, peak activity had an approximate molecular weight of 53,000 and appeared distinct from recognized forms of lymphotoxin.

Amino Acids↗
PubMedDate not supplied

Detection of aldehyde dehydrogenase deficiency in Chachi Indians, Ecuador.

The activity of mitochondrial aldehyde dehydrogenase (ALDH2) was tested by isoelectric focusing of hair root extracts from 50 Chachi Indians (Ecuador). Quality of extracts and the intactness of cytoplasmic and mitochondrial enzymes were ascertained by assaying of phosphoglucomutase (PGM) and malate dehydrogenase (MDH) in the same extracts. Three of the 39 successfully assayed Chachi Indian samples showed virtual absence of the ALDH2 band on the isoelectropherogram, and the control enzymes were stained normally in these subjects. These data confirm the existence of a mitochondrial ALDH deficiency among South American Indians. The molecular origin of the ALDH2 deficiency in this population is unknown.

Adolescent↗
PubMedDate not supplied

Wuchereria bancrofti: cloning and characterization of heat shock protein 70 from the human lymphatic filarial parasite.

Heat shock protein 70 (HSP70) was identified as an immunodominant antigen by screening a Wuchereria bancrofti (Wb) microfilarial cDNA library with pooled Wb-infected sera, with 28% of the immunopositive clones coding for Wb-HSP70. The deduced amino acid sequence showed greater than 97 and 85% identity with HSP70 from filarial nematodes and humans, respectively. Recombinant HSP70 (74 kDa) and a recombinant protein from the C-terminal portion (43 kDa) also reacted with pooled Wb-infected sera, suggesting that the C-terminal region of HSP70 contains at least one antibody epitope. Brugia malayi L3 larvae showed increasing levels of HSP70 with increasing temperatures. Further, a polyclonal mouse anti-Wb-HSP70 antibody had reactivity to the HSP70 of cattle filarial parasite Settaria digitata and to human HSP70 derived from a Hep-2 cell line. Immune reactivity to Wb-HSP70 was strong, with uninfected non-endemic normal sera showing significantly greater reactions than sera from filaria-infected individuals. Both immunodominant self-HSP70 and HSP70 from other microbial infections may be primary targets for developing autoantibodies naturally.

Amino Acid Sequence↗