Gastrointestinal candidiasis in a murine model of severe combined immunodeficiency syndrome.

A murine model of severe combined immunodeficiency syndrome (scid mice) affords an opportunity to study the interaction of Candida albicans with a host lacking functional B- and T-cell mechanisms. We have previously reported no significant difference in yeast recovery after intravenous challenge of BALB/c mice and scid mice with C. albicans (S. Mahanty, R.A. Greenfield, W.A. Joyce, and P.W. Kincade, Infect. Immun. 56:3162-3166, 1988). In this study, we evaluate the course of gastrointestinal candidiasis after a single oral challenge with C. albicans. BALB/c and scid mice received H2O containing 10(6) C. albicans per ml for 16 h. Half the mice of each strain continuously received H2O containing 1 mg of tetracycline per ml. Stool samples were cultured for yeast twice weekly until they were negative three consecutive times or positive for 8 weeks. Mice were then sacrificed for quantitative cultures of liver, spleen, and kidneys. At eight weeks postinoculation, 2 of 13 BALB/c mice, 0 of 14 BALB/c mice receiving tetracycline, 6 of 12 scid mice, and 8 of 13 scid mice receiving tetracycline had positive stool cultures (P less than 0.05, likelihood ratio chi-square). Quantitative recovery of yeasts from stools was also higher in the scid mice. Cultures of liver, spleen, and kidneys wer negative in all BALB/c mice and essentially all negative in scid mice; a single colony was isolated from the kidney of one scid mouse and the liver of another scid mouse. We conclude that B cells and/or T cells and their products are important in gastrointestinal colonization with C. albicans but that even in their absence, dissemination of infection from the gastrointestinal tract does not consistently occur. Thus, other aspects of host defense must be critical in containing gastrointestinal Candida colonization.     

Differences in predominant T cell phenotypes and distribution pattern in reactional lesions of tuberculoid and lepromatous leprosy.

The nature and histological pattern of the cutaneous infiltrates of 17 leprosy patients in reversal reactions (Type I) and erythema nodosum leprosum (Type II, ENL) were compared with tissues from 18 non-reactional borderline leprosy (BT, BL) and lepromatous leprosy (LL) patients using monoclonal antibodies and immunofluorescence. Reactional BT lesions showed a mild increase in OKT11+ pan T cells as compared to non-reactional tissues and a significant influx of OKT8+ (suppressor/cytotoxic) cells which were peripherally localized in the lymphocyte mantle surrounding the epithelioid cells. The Leu 3a+ (helper/inducer) cells were scattered amongst the lymphocytes and macrophages. The mean ratio (+/- s.d.) of Leu 3a+/OKT8+ cells was 1.88 +/- 0.64 in Type I BT reactions as compared to 2.95 +/- 0.95 in BT lesions. In contrast, lesions of BL reversal reactions and ENL showed a more marked increase in pan T cells with a preponderance of the helper/inducer subset, Leu 3a+/OKT8+ ratio being 2.26 +/- 0.61 and 0.93 +/- 0.57 in BL reactional and non-reactional lesions, respectively. Interestingly, this increase in the numbers of the T cells reached levels observed in BT lesions. The distribution pattern of OKT8+ cells was similar to Leu 3a+, both being diffusely scattered amongst the bacilli laden macrophages. Ia like antigens were present in all granulomas and were abundant on lymphocytes and macrophages and less conspicuous on epithelioid cells. T6+ Langerhans cells were uniformly increased in all reactional lesions. It would appear that the changes observed in both Type I and Type II reactions are similar in the lepromatous group of patients. They differ significantly from the BT reversal reaction in terms of the dominant T cell subset and the microanatomical distribution of the OKT8+ cells in the lesions.     

Immunohistologic comparison between armadillo-derived leprosin and standard lepromin skin tests in leprosy patients

A comparison was made on the in situ immunological characteristics of dermal infiltrates of early (24-hour) and late (3-4 weeks) skin reactions in leprosy patients. The skin reactions were induced by armadillo-derived leprosin coupled to liposomes and standard Dharmendra lepromin. Most lymphocytes in the early reaction induced by both antigens were positive for Leu 4, Leu 3a, OKT8 and Ia like antigens indicating thereby the presence of activated T cells. The ratio of Leu 3a/OKT8+ cells were similar. In the late reaction elicited by both antigens, the lymphocytes in the granulomas were predominantly activated T lymphocytes expressing Leu 4, Leu 3a, OKT8 and Ia like antigens. Leu 3a+ cells were scattered diffusely amidst the epithelioid cells. In contrast, the OKT8+ cells were present mainly as 'a ring' in the periphery of the granuloma. A similar ratio of Leu 3a+/OKT8+ cells was observed in these granulomas. Macrophages in the granulomas expressed Ia like antigens. These observations indicate that the immunological characteristics of dermal infiltrates in the skin reaction induced by armadillo-derived leprosin coupled to liposomes and standard Dharmendra lepromin appear to be identical.     

Bain type of X‐linked syndromic mental retardation in a male with a pathogenic variant in HNRNPH2

Heterogeneous nuclear ribonucleoproteins (hnRNPs) are RNA binding proteins, which aid in maturation, stabilization, and transport of mRNA. They have a significant role in cellular nucleic acid metabolism. The hnRNPs alter gene expression and are linked to various neurodegenerative disorders and cancers. Previously, six unrelated girls with developmental delay, intellectual disability, and hypotonia were found to have de novo heterozygous pathogenic missense variants in HNRNPH2, located on the X chromosome. A gain‐of‐function effect was proposed for the variant and it was thought to be lethal in males as no surviving males were identified. We describe a family with two affected siblings, one male and one female, with a known pathogenic variant in HNRNPH2, possibly due to maternal germline mosaicism.     

In situ demonstration of Mycobacterium leprae antigens in leprosy lesions using monoclonal antibodies

Cryostat sections of skin and nerve lesions of leprosy were stained with monoclonal antibodies recognising Mycobacterium leprae antigens and indirect immunofluorescence. In both the tuberculoid and lepromatous lesions, PGL1, 55-65-kDa, 17-kDa protein antigens and cross-reactive non-protein antigens were present. 65-kDa antigens were seen mainly in the skin lesions of lepromatous leprosy. The infiltrates in both the skin and nerve granulomas of tuberculoid and lepromatous leprosy showed membranous staining with monoclonal antibodies recognising PGL1 and 55-65-kDa antigens. Bacilli in the lesions and the cells in the lymph node granulomas of patients with tuberculosis or the infiltrates in the lesions of tinea corporis or sections of normal skin did not show any staining with these monoclonal antibodies. These results confirm that M. leprae antigens are present and are expressed on the infiltrating cells of leprosy lesions.     

Processing sputum specimens in a refrigerated centrifuge does not increase the rate of isolation of Mycobacterium tuberculosis

A total of 1,047 sputum samples from pulmonary tuberculosis patients was collected in cetyl pyridinium chloride-sodium chloride solution. Each sample was divided into two parts and randomly allocated for the isolation of Mycobacterium tuberculosis, with one part to be processed by the standard method and the other by a modified method. In the standard method, the samples were processed by using nonrefrigerated centrifuges, while in the modified method, they were processed by using a refrigerated centrifuge. Fifty-seven samples that yielded contaminants were excluded, and the remaining 990 samples were taken up for analysis. The rates of isolation of M. tuberculosis with the standard and modified methods were 48.6 and 48.1%, respectively, and the difference was not statistically significant (McNemar's test; P > 0.5). However, 51% of the positive cultures were isolated within 2 weeks with the modified method compared to 37% with the standard method (chi-square test; P < 0.001). The results of the study reveal that processing of sputum samples in a refrigerated centrifuge does not improve the rate of isolation but will result in rapid isolation of M. tuberculosis.     

Molecular characterization of a calcium binding translationally controlled tumor protein homologue from the filarial parasites Brugia malayi and Wuchereria bancrofti

We have cloned homologues of the mammalian translationally controlled tumor protein (TCTP) from the human filarial parasites Wuchereria bancrofti and Brugia malayi. TCTP genes from B. malayi and W. bancrofti were expressed in a T7 promoter vector as histidine tagged fusion proteins. Both the recombinant B. malayi TCTP (rBm-TCTP) and recombinant W. bancrofti TCTP (rWb-TCTP) have a molecular mass of approximately 28 kDa with the histidine tag. Sequence analyses showed that there is a 98% similarity between the two filarial TCTPs at amino acid levels and are immunologically cross-reactive. Analysis of soluble proteins from various lifecycle stages of B. malayi suggested that the expression of Bm-TCTP might be differentially regulated and occurs in multimeric form. Recombinant TCTP were found to form multimers in solution under non-reducing conditions. The tendency for filarial TCTPs to become multimers was predicted by the presence of the Lupas coiled coil structure in their sequence. Despite the absence of a signal sequence, Bm-TCTP is present abundantly in the excretory/secretions (ES) of microfilariae. Characterization studies showed that both Bm- and Wb-TCTPs are calcium-binding proteins and have histamine-releasing function in vitro. When injected intraperitoneally both the filarial TCTPs induced inflammatory infiltration of eosinophils into the peritoneal cavity of mice suggesting that the filarial TCTPs may have a role in the allergic inflammatory responses associated with filarial infections.     

Bone morphogenetic protein 2 induces the expression of cementum attachment protein in human periodontal ligament clones

Cementum is continuously formed during the lifetime of a tooth. The paravascular zones in the adult periodontal ligament (PL) comprise the progenitors for the fibroblastic (Fb) lineage and mineralized tissue-forming (MTF) cell lineages--the osteoblastic (Ob) and cementoblastic (Cb) lineages. Recent studies indicate that cementum attachment protein (CAP) is related to the differentiation of the Cb lineage and is instrumental in differentiating between the three periodontal cell lineages. The purpose of this study was to assess the effect of bone morphogenetic protein 2 (BMP2) on the expression of cementum attachment protein (CAP) and on the differentiation of cloned PL progenitors. The effect of BMP2 on CAP expression and on the differentiation of cloned Fb and MTF progenitors was tested by assessing the expression of alkaline phosphatase (ALP), CAP, and bone sialoprotein (BSP) by immunochemistry and by determining the CAP-binding capacity of these clones. Untreated Fb clones were negative for all tested markers and had low CAP-binding capacity. Untreated MTF clones had a high CAP-binding capacity and were positive for the three markers. BMP2 enhanced the CAP-binding potential of both Fb and MTF clones. BMP2 induced the expression of CAP, ALP, and BSP in the Fb clones and enhanced the expression of CAP and BSP in the MTF clones. These results indicate for the first time that BMP2 can recruit progenitors to the Cb lineage and regulate the differentiation of the Cb lineage by inducing and enhancing the expression of CAP, a cell lineage-specific regulator. Furthermore, the results suggest that the MTF and Fb lineages may originate from a common early progenitor cell.     

Depression of T-cell function and normality of B-cell response in protein calorie malnutrition.

Antibody response to a T-cell dependent antigen, sheep erythrocytes and to a B-cell mitogen, purified lipopolysaccharide (LPS), has been studied in mice kept on protein deficient (2 and 4 per cent casein) diets. The number of plaque-forming cells (PFC) to SRBC were 20-5 +/- 7-7 per million spleen cells in protein-deficient animals compared to 261-0 +/- 31-1 in parallel controls maintained on a protein rich diet (18 per cent casein). No difference was observed in number of PFC formed in controls and deficient animals to LPS, values were 161-4 +/- 19-7, 158-5 +/- 14-2, & 162-3 +/- 31-9 in control (18 per cent casein) and deficient groups (4 per cent and 2 per cent casein) respectively. The delayed hypersensitivity skin reaction to SRBC measured in foot pads was significantly lower in mice on 4 per cent casein diet compared to controls. These studies suggest that the effect of protein deficiency is primarily on T-cell function and not on the B-cell response;