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Biomedical subjects

J J Castles

Publications and source records attributed to J J Castles.

At least 19 recordsLinked to original sources
PubMed1988

Localization, molecular weight and immunoglobulin subclass response to Aspergillus fumigatus allergens in acute bronchopulmonary aspergillosis.

A specimen of Aspergillus fumigatus was isolated from a patient with acute bronchopulmonary aspergillosis (ABPA). Cultures were allowed to propagate and were separated into spores and mycelium to greater than 95% homogeneity. Extracts of both the spores and mycelium were prepared and used for study by both dot blot analysis and immunoblotting to study the major allergens. By dot blot analysis, 22 of 22 patients with ABPA and none of 10 healthy controls reacted with mycelium when probed for both IgG and IgE anti-Aspergillus reactivity. Similar results were obtained when these extracts were separated on polyacrylamide gel electrophoresis and then probed; this included both IgG and IgE reactivity. Further, the reactivity to preparations of whole extract was absorbed with mycelium but not spores. Several distinct protein bands were detected with IgG, ranging from 30 to 110 kilodaltons (kD). In contrast, the majority of patients with ABPA, when studied for IgE reactivity, reacted only with a 70-kD protein, although 1 out of 22 patients reacted with a 30-kD protein. Subclass analysis of IgG reactivity demonstrated that 90% of reactive sera contained IgG2 and 73% IgG4 reactivity. IgG1 and IgG3 reactivity were present but less frequently detected. The use of immunoblotting will enable the identification of relevant allergens. Further, the demonstration that the allergenicity is located primarily in mycelium will allow more definitive studies of allergen isolation for studies of reactivity and, ultimately, cloning of the 70-kD protein and, finally, epitope mapping.

Acute Disease↗
PubMed1987

Abnormalities of B lineage cells are demonstrable in long term lymphoid bone marrow cultures of New Zealand black mice.

The formation of B lymphocytes in young New Zealand Black (NZB) mice proceeds at an accelerated rate resulting in a deficiency of B lineage precursors in adult (greater than 15 wk old) animals. To study the characteristics of B lineage cells in young (4 wk) and old (6 mo) NZB mice, bone marrow from these animals was used to initiate long term lymphoid bone marrow cultures (LBMC) that permit the long term maintenance of B cells and their precursors. Age-matched cultures from BALB/c mice and NZB.xid marrow were established in parallel. Primary LBMC were readily established from these strains and showed similar patterns of growth for the 3-mo observation period. No significant differences in numbers of 14.8 positive cells were observed. However, NZB mice at both ages had a higher percentage of membrane IgM (mIgM)-expressing cells. Significant levels of supernatant IgM were found only in cultures of 6-mo NZB and BALB/c mice; levels were highest in NZB culture supernatants and were often more than 500 ng/ml; significant, although much lower, levels of IgG were likewise detected. Lymphoid cells from NZB.xid mice were unable to generate significant levels of IgM in supernatant fluids indicating the effects of the xid gene were displayed in vitro. Autoantibodies were not detected in any of the culture supernatants. Additional evidence for NZB hyperactivity in primary B lymphopoiesis was observed upon initiation of primary myeloid bone marrow cultures (MBMC) from these strains of mice and subsequently transferring them to LBMC conditions. This results in the cessation of myelopoiesis at the initiation of B lymphopoiesis. At the time of converting MBMC to LBMC, cultures of NZB and BALB/c mice morphologically resembled myeloid cultures and had neither B cell colony-forming units nor cells that expressed 14.8 or mIgM. However, following the switch, NZB mice had a 5-fold higher number of B cell colony-forming units. Further, MBMC established from NZB bone marrow cells had a reduced capacity to form colonies in the granulocyte-macrophage colony-forming unit assay. These studies indicate that defects of NZB hemopoietic cells are manifest in vitro and suggest the use of in vitro long term cultures as a valuable technique to further dissect the hematopoietic abnormalities of NZB mice and possible underlying microenvironmental defects.

Animals↗
PubMed1987

The use of congenital immunologic mutants to probe autoimmune disease in New Zealand mice.

To further our understanding of autoimmunity, many laboratories have concentrated on the study and manipulation of murine lupus in several strains. Although direct extrapolation of data from animal to man must proceed with caution, the use of such animal systems, both in vitro and in vivo, has been an enormous help in the development of immunologic concepts. Our laboratory has been studying murine lupus by selective breeding of specific genetic immune defects onto New Zealand mice. Specifically, we have used congenital immunologic mutations resulting in asplenia (Dh/+), athymia (nu/nu) and immunodeficiency (Xid) as a means of probing the natural history of immunopathology in one such murine model of autoimmunity New Zealand (NZ) mice. These studies have provided important insights into the ontogeny of autoimmunity. NZB.Xid mice have been particularly valuable and have become a useful tool for dissecting the B cell defects of NZ mice. The Xid gene is dominant over the premature polyclonal activation of NZB mice and acts almost exclusively on B cells that are involved in autoantibody production in NZB mice. Nonetheless, the fact that autoantibody production can occur in a small percent of very old NZB.Xid mice, which have the same phenotype as other NZB.Xid mice, suggests that it can be produced by a mechanism other than a generalized polyclonal expansion and is not dependent on the circulatory or splenic frequency of the Lyb 5 subset of cells. Finally, NZB.Xid mice are unable to produce autoantibodies even after maturing in an aged NZB microenvironment, which suggests that the cell population missing in NZB.Xid mice are important for autoantibody production.

Animals↗
PubMed1985

Tissue localization and biochemical characteristics of a new thymic antigen recognized by a monoclonal thymocytotoxic autoantibody from New Zealand black mice.

Naturally occurring thymocytotoxic autoantibodies (NTA) have been described in both humans and mice with SLE. To define further the role of anti-thymic autoantibodies in murine lupus, we studied the cellular and molecular specificity of a spontaneous monoclonal NTA, designated TC-17, derived from a 4-mo-old New Zealand Black mouse. TC-17, an IgM autoantibody, has been shown previously to be unreactive with Lyt-1, Lyt-2, and L3T4 (T helper) antigens. We have shown further that it is also unreactive with Thy-1. TC-17 recognizes a new thymic antigen that appears to mark a distinct subpopulation of cortisol-sensitive cortical thymocytes. The antigen consists of a single glycoprotein chain with an apparent m.w. of 88,000. TC-17 shows reduced binding to thymocytes treated with tunicamycin, indicating either that glycosylation of TC-17 antigen is necessary for TC-17 to bind to it or that glycosylation is required for expression of the antigen on the cell surface. TC-17 uniquely reacts with two of 17 murine lymphoid tumor cell lines of intermediate cellular maturity. The thymocytotoxic activity of TC-17 is absorbed by single cell suspensions of murine stomach, small intestine, large intestine, kidney, and thymus. Moreover, the specific binding of TC-17 to gut tissue of normal and germfree mice can be demonstrated by indirect immunofluorescence, suggesting antigenic cross-reactions between thymic and gut tissue. TC-17 reacts with rat thymocytes as well as it does with murine cells, indicating moderate evolutionary conservation of the TC-17 antigen. The expression of this glycoprotein by a discrete thymocyte subset may prove to be a valuable probe for the study of murine T cell differentiation.

Absorption↗
PubMed1984

Induction of erythrocyte autoantibodies in NZB mice: spectrotype and relationship with the Xid gene.

The production of autoantibodies to erythrocytes by immunization with rat red blood cells (RRBC) is significantly retarded in X-linked immune deficient (Xid) mice. We have attempted to further explore this relationship by characterizing RRBC-induced erythrocyte autoantibodies in high responder New Zealand Black (NZB) and congenic NZB.Xid mice. NZB.Xid animals, immunized with RRBC, readily produce anti-RRBC antibodies and cross-reactive antiautologous erythrocyte antibody (CR anti-MRBC) as well as anti-HB antibodies. The autoantibody response of NZB.Xid mice to RRBC appears similar to NZB controls with respect to both the time of onset and subclass diversity; the anti-HB antibody cannot be absorbed with RRBC. Moreover, there are no alterations in the spectrotype of antierythrocyte antibodies found in NZB.Xid mice. Nonetheless, NZB.Xid, but not NZB mice, fail to produce splenic plaque-forming responses against bromelase-treated mouse red blood cells. Lyb 5+ cells are not required for the production of RRBC-induced antierythrocyte autoantibodies. These results, when discussed in light of the low but significant incidence of spontaneous DAT in NZB.Xid mice, suggest that given the appropriate genetic repertoire, the influence of the Xid gene on autoantibody production can be bypassed.

Anemia, Hemolytic, Autoimmune↗
PubMed1983

Anti-mu induces lymphoma in germfree congenitally athymic (nude) but not in heterozygous (nu/+) mice.

To further our understanding of the role of host immunity in the development of lymphoid neoplasia, groups of germfree BALB/c nude and nu/+ mice were either followed unmanipulated or were treated, beginning at birth, with anti-mu, normal goat IgG or with lipopolysaccharide (LPS). The survival and development of neoplasia of all groups of animals were monitored up to 2 yr. Nude mice, under germfree and specific pathogen-free (spf) conditions, had a higher incidence of lymphoid neoplasia and reduced survival when compared to nu/+ littermates. The incidence of lymphoid tumors in nude mice under spf or germfree conditions was 7.2 and 8.7%, respectively, in comparison to 0% in nu/+ animals. Treatment of germfree nude mice with anti-mu, but not with goat IgG, increased the incidence of lymphoid tumors to 39%. Anti-mu did not significantly change the incidence of lymphoid neoplasia in nu/+ animals. Treatment of nu/nu and nu/+ mice with LPS, however, led to a several-fold increase in the appearance of neoplasia, to values of 25.4% in nude and 10% in nu/+ mice. Lymphoid neoplasia found in either unmanipulated, anti-mu, or IgG-treated germfree or spf mice included Thy-1.2+, surface IgM+, and IgG+ tumors. In contrast, all the lymphomas found in LPS-treated mice were surface IgM+. Thus, whereas LPS may have generated a relatively homogeneous group of tumors, anti-mu may have randomly increased the normal incidence of spontaneous tumors. Moreover, although there was significant variation in the histologic appearance of tumors, both within treatment groups as well as in different areas of the same animal, only LPS-treated mice were regularly noted to have distant nonlymphoid involvement, with lesions found in liver, lung, and kidney. In contrast, the incidence of nonlymphoid neoplasia was similar and was less than 2.5% in all groups.

Aging↗
PubMed1981

Characterization of a spontaneous disease of white leghorn chickens resembling progressive systemic sclerosis (scleroderma).

University of California, Davis (UCD) line 200 White Leghorn Chickens spontaneously develop a syndrome that has many analogous features to human progressive systemic sclerosis. This syndrome is characterized by progressive involution of comb, dermal fibrosis, and distal polyarthritis. These three features occur within 6 wk after hatching, and are accompanied by a 40% mortality as a result of vaso-occlusive disease, with development of secondary infection of peripheral gangrenous lesions. Birds that survive greater than 2 mo after hatching progressively develop fibrosis of the esophagous and mononuclear infiltration of heart and kidney, with prominent occlusion of small and medium sized blood vessels. In addition, line 200 chickens develop rheumatoid factors, antinuclear antibodies, and antibodies to collagen, but do not have antibodies to thymocytes, DNA, or extractable nuclear antigens. Moreover, antinuclear antibodies when studied using HEp-2 cells as substrate demonstrate predominantly a speckled pattern. This syndrome of line 200 chickens is not detectable in F1 crosses to several UCD inbred lines. F1 X parental line BC1 backcrosses have an approximately 50% incidence of disease, suggesting that this syndrome is inherited as autosomal recessive. However, only 4% of F2 generation birds show abnormal symptoms, suggesting the presence of modifying genes. There is no appearance of IgG deposition, as determined by immunofluorescence, in either skin, blood vessels, esophagus, or heart. However, approximately 20% of chickens have a glomerulonephritis; this feature appears to be a terminal event and does not appear clinically significant. Although this syndrome of line 200 chickens has several features that are in sharp distinction to human scleroderma, the presence of common immunologic and pathologic denominators suggest that this spontaneous disease may be an appropriate model to develop a better understanding of autoimmune connective tissue diseases.

Age Factors↗
PubMed1980

Accelerated plasmacytoma formation in mice treated with alpha-fetoprotein.

The ability of alpha-fetoprotein (AFP), derived from mouse amniotic fluid, to alter qualitative and quantitative characteristics of pristane-induced plasmacytomas and the carcinogenic action of 3-methylcholanthrene (MCA) and 7,12-dimethylbenz]a[-anthracene (DMBA) was studied and compared to analogous treatment with murine albumin and transferrin in BALB/c mice. Pharmacologic doses of 200 microgram AFP ip three times per week had no effect on either latency period or tumor incidence in mice given injections of MCA and DMBA when compared to albumin-treated and transferrin-treated controls. Moreover, this lack of influence of AFP was found at high and low dosages of the carcinogens. In contrast AFP, but neither albumin nor transferrin, accelerated the appearance of plasmacytomas in pristane-primed BALB/c mice. Moreover, the immunoglobulin class of plasmacytomas of mice given AFP injections was qualitatively altered, and in increase in IgM and IgG2b plasmacytomas was noted. These effects occurred at serum AFP levels of 90--260 microgram/ml. Although these results should not be used to support a normal physiologic role of AFP in immune regulation, they are similar to previous observations of suppression mediated by the alpha-globulin fraction of normal serum and suggest that pharmacologic dosages of AFP may be useful in the study of select, possibly thymus-dependent, immune responses.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1979

Studies of congenitally immunologic mutant New Zealand mice. II. Absence of T cell progenitor populations and B cell defects of congenitally athymic (nude) New Zealand Black (NZB) mice.

Congenitally athymic (nude) mice on an NZB, NZW, and BALB/c background were produced by repetitive selective backcrossing. F'12 generation nude mice of these three strains were compared to their littermate nu/+ controls with respect to survival, histology, blood counts, splenic surface markers, response to mitogens, spontaneous plaque-forming cells, and appearance of naturally occurring thymocytotoxic antibodies (NTA). Under specific pathogen-free conditions, NZB nude mice survive less than 3 weeks, dying of a runting-like disease with infection by local normally noninvasive organisms. A contributing factor to his premature death is the relative absence of T cell progenitor populations in the NZB nude vs NZW nude or BALB/c nude groups. Furthermore, NZB nude mice have a significantly earlier appearance of NTA than nu/+ littermates and likewise appear to have heightened spontaneous polyclonal B cell responses against the haptens dansyl, nitroiodophenyl, trinitrophenyl,2,4 dinitrophenyl, and sulfonate. It is suggested that NZB mice have several critical immunologic defects, including abnormalities of thymic epithelial cells, T cell differentiation pathways, and chronically polyclonal activated B cell populations. These defects interact to produce the clinical expression of autoimmunity.

Animals↗
PubMed1979

Altered responsiveness to mitogens in subgroups of patients with juvenile rheumatoid arthritis.

Thirty patients with juvenile rheumatoid arthritis (JRA) were compared to an age-matched control population with respect to lymphocyte transformation by the mitogens Concanavalin A, Phytohemagglutinin-P, Pokeweed and Lipopolysaccharide, as well as for the relative frequency of E-rosette and Ig-bearing cells in peripheral blood. The data were analysed according to sex, age of onset, mode of onset and disease activity. Lymphocytes from patients with a pauciarticular pattern of onset of JRA responded to mitogens in a similar fashion as normal volunteers, whereas lymphocytes from patients with both a polyarticular and Still's mode of onset had a diminished response to the mitogens. However, this reduction was noteworthy only during periods of disease activity; during periods of remission, lymphocytes from the patients responded similarly to lymphocytes from normals. In contrast to this reduction in mitogen stimulation in these subgroups of patients with JRA, all patients with JRA--irrespective of disease activity or mode of onset--were similar to controls with respect to the percentage of E-rosette and Ig-bearing cells. These alterations in mitogen responsiveness could not be attributed to therapy. Finally, the abnormalities of lymphocyte transformation in patients with polyarticular and Still's mode of onset represent secondary features of disease and are not due to a generalized predisposition or abnormality of cell mediated immunity.

Adolescent↗