T lymphocytes and acute kidney injury: update.

The immune system is among the key pathogenic factors in acute kidney injury (AKI). Various immune cells, including dendritic cells, natural killer T cells, T and B lymphocytes, neutrophils and macrophages are involved. Conventional CD4+ lymphocytes are well established to participate in early injury, and CD4+CD25+FoxP3 regulatory T cells are protective and can accelerate repair. A newly identified kidney T cell receptor + CD4-CD8- (double-negative) T cell has complex functions, including potentially anti-inflammatory roles in AKI. In this mini review, we summarize the data on the role of lymphocytes in AKI and set the stage for further mechanistic studies as well as interventions to improve outcomes.

High-fat diet modulates non-CD1d-restricted natural killer T cells and regulatory T cells in mouse colon and exacerbates experimental colitis.

Environmental factors such as diet are known to play important roles in inflammatory bowel disease (IBD). Epidemiological studies have indicated that a high-fat diet is a risk factor for IBD. In addition, the balance between effector T cells (T(eff)) and regulatory T cells (T(reg)) contributes to the pathogenesis of mucosal inflammation. The aim of this study was to understand the mechanisms by which a high-fat diet can regulate susceptibility to intestinal inflammation. Wild-type C57BL/6 mice were fed either a commercial high-fat diet or a normal diet, then exposed to dextran sulphate sodium (DSS) to induce colonic inflammation. Intraepithelial lymphocytes (IEL) were isolated from the colon, and their phenotype and cytokine profile were analysed by flow cytometry. Mice receiving the high-fat diet were more susceptible to DSS-induced colitis. They had higher numbers of non-CD1d-restricted natural killer (NK) T cells in the colonic IEL, when compared to mice fed a normal diet. These cells expressed tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma, which are up-regulated by high-fat diets. Mice fed the high-fat diet also had decreased levels of colonic T(reg). Depletion of colonic NK T cells or adoptive transfer of T(reg) reduced the DSS colitis in these mice, and reduced the colonic expression of TNF-alpha and IFN-gamma. We conclude that a high-fat diet can increase non-CD1d-restricted NK T cells and decrease T(reg) in the colonic IEL population. This altered colonic IEL population leads to increased susceptibility to DSS-induced colitis. This effect may help to explain how environmental factors can increase the susceptibility to IBD.

Brucellosis among animals and human contacts in eastern Sudan.

OBJECTIVE: To study the relative frequency of brucellosis among domestic animals in Kassala State, Sudan, in the year 1999 and compare the results of previous studies since 1908. Also to study the frequency rates of the disease in animal contacts in the area. METHODS: Sero-survey for Brucella melitensis and Brucella abortus was carried out on sera of animals brought for slaughter to Kassala abattoir and on sera of occupational contacts of animals. All sera were tested by the slide agglutination test. The positive reactors were confirmed by tube agglutination test. RESULTS: A total of 1225 sera were tested - 1038 were animal sera and 187 were human sera. Four percent of goats sera, 1% of sheep sera and 5% of cattle sera were found to be positive. Of the 64 camel sera tested, none were positive reactors. Of the occupational contacts, which included butchers, slaughterhouse workers, milkers and cow attendants, (1%) reacted positively. CONCLUSION: The study showed low frequency rates of brucellosis among animals in the Kassala area compared with other parts of Sudan. Occupational contacts showed very low frequency rates of the disease. This is often over-looked by medical practitioners due to the overwhelming problem of malaria in the area. We draw the attention of practitioners to think of brucellosis in the differential diagnosis of fever especially in rural communities.

Lack of coreceptor allows survival of chronically stimulated double-negative alpha/beta T cells: implications for autoimmunity.

Lymphoproliferative diseases are characterized by massive accumulation of CD4(-)CD8(-)B220(+) (double-negative [DN]) T cells in peripheral organs. Although evidence indicates these cells are derived from mature autoreactive alpha/beta T cells, the significance of coreceptor downregulation is not known. In this study, we examined the role CD4 coreceptor plays in the survival of repeatedly stimulated T cells. CD4(+/+) and CD4(-/-) T cells from AND T cell receptor (TCR) transgenic mice exhibited similar phenotypes after antigenic stimulation, but the CD4(-/-) T cells survived in much larger numbers than the CD4(+/+) cells upon primary and secondary major histocompatibility complex (MHC)/peptide stimulation. Enhanced survival of CD4(-/-) T cells was due to decreased apoptosis rather than enhanced proliferation. Similarly, circumvention of the CD4/MHC interaction by using a surrogate TCR ligand that does not engage CD4 led to significant enhancement of CD4(+/+) cells than when stimulated with MHC/peptide. Finally, we generated DN B220(+) T cells using an in vitro model system and showed they were more tolerant to chronic stimulation than CD4(+/+) cells. Together, these results indicate that coreceptor engagement controls expansion of normal T cells. In the absence of coreceptor, T cells survive chronic stimulation and express B220 as seen in autoimmune lymphoproliferative diseases.

Antigen-induced T cell death is regulated by CD4 expression.

Activation induced cell death (AICD) is a major physiologic pathway that regulates T cell homeostasis. In CD4 T cells, AICD is mediated mainly through Fas/FasL interactions. Although TCR occupancy triggers AICD, the contribution of its tightly associated CD4 coreceptor to the process that leads to AICD is not known. Here we show that CD4 molecule plays an essential regulatory role of TCR dependent AICD. Loss of CD4 rendered activated 5kc T cell hybridoma resistant to AICD. The resistance of CD4 negative 5kc T cells to AICD was due to selective inhibition of FasL expression and it could be reversed by addition of recombinant FasL. Furthermore, a direct functional link between CD4 and FasL was demonstrated by induction of FasL upon CD4 crosslinking in a TCR independent fashion. The importance of CD4 interaction with MHC/peptide complex in mediating AICD was also evident in normal T cells that could survive chronic stimulation with anti-CD3 but died after short period of proliferation after stimulation with MHC/peptide. Thus it appears that AICD is controlled by the CD4 molecule via regulation of FasL expression. These findings have important implications for our understanding of mechanisms of peripheral tolerance as well as pathogenesis of autoimmune diseases.

Soluble, high-affinity dimers of T-cell receptors and class II major histocompatibility complexes: biochemical probes for analysis and modulation of immune responses.

T cell receptors (TCR) and major histocompatibility complex (MHC) molecules are integral membrane proteins that have central roles in cell-mediated immune recognition. Therefore, soluble analogs of these molecules would be useful for analyzing and possibly modulating antigen-specific immune responses. However, due to the intrinsic low-affinity and inherent solubility problems, it has been difficult to produce soluble high-affinity analogs of TCR and class II MHC molecules. This report describes a general approach which solves this intrinsic low-affinity by constructing soluble divalent analogs using IgG as a molecular scaffold. The divalent nature of the complexes increases the avidity of the chimeric molecules for cognate ligands. The generality of this approach was studied by making soluble divalent analogs of two different classes of proteins, a TCR (2C TCR2Ig) and a class II MHC (MCCI-Ek2Ig) molecule. Direct flow cytometry assays demonstrate that the divalent 2C TCR2Ig chimera retained the specificity of the native 2C TCR, while displaying increased avidity for cognate peptide/MHC ligands, resulting in a high-affinity probe capable of detecting interactions that heretofore have only been detected using surface plasmon resonance. TCR2IgG was also used in immunofluorescence studies to show ER localization of intracellular peptide-MHC complexes after peptide feeding. MCCI-Ek2Ig chimeras were able to both stain and activate an MCC-specific T cell hybridoma. Construction and expression of these two diverse heterodimers demonstrate the generality of this approach. Furthermore, the increased avidity of these soluble divalent proteins makes these chimeric molecules potentially useful in clinical settings for probing and modulating in vivo cellular responses.

Potent T cell activation with dimeric peptide-major histocompatibility complex class II ligand: the role of CD4 coreceptor.

The interaction of the T cell receptor (TCR) with its cognate peptide-major histocompatibility complex (MHC) on the surface of antigen presenting cells (APCs) is a primary event during T cell activation. Here we used a dimeric IEk-MCC molecule to study its capacity to activate antigen-specific T cells and to directly analyze the role of CD4 in physically stabilizing the TCR-MHC interaction. Dimeric IEk-MCC stably binds to specific T cells. In addition, immobilized dimeric IEk-MCC can induce TCR downregulation and activate antigen-specific T cells more efficiently than anti-CD3. The potency of the dimeric IEk-MCC is significantly enhanced in the presence of CD4. However, CD4 does not play any significant role in stabilizing peptide-MHC-TCR interactions as it fails to enhance binding of IEk-MCC to specific T cells or influence peptide-MHC-TCR dissociation rate or TCR downregulation. Moreover, these results indicate that dimerization of peptide-MHC class II using an IgG molecular scaffold significantly increases its binding avidity leading to an enhancement of its stimulatory capacity while maintaining the physiological properties of cognate peptide-MHC complex. These peptide-MHC-IgG chimeras may, therefore, provide a novel approach to modulate antigen-specific T cell responses both in vitro and in vivo.

Transcytosis of staphylococcal superantigen toxins.

Staphylococcus aureus produces a set of proteins (e.g., staphylococcal enterotoxin A [SEA], SEB, toxic shock syndrome toxin 1 [TSST-1]) which act both as superantigens (SAgs) and toxins. Although their mode of action as SAgs is well understood, little is known about how they enter the body via the intestine and cause food poisoning. To examine this problem we used an in vitro culture system to study the capacity of class II MHC-negative human intestinal epithelial cells (Caco-2) to transcytose several staphylococcal toxins. We found that Caco-2 cells are capable of dose-dependent, facilitated transcytosis of SEB and TSST-1, but not SEA. We extended these studies in vivo in mice by showing that ingested SEB appears in the blood more efficiently than SEA. Our data suggest that these toxins can cross the epithelium in an immunologically intact form. These results may have important implications for the pathogenesis of food poisoning.

Murine transfer factors: dose-response relationships and routes of administration.

Transfer factors are protein immunomodulators that transfer the ability to express cell-mediated immunity from immunized donors to nonimmune recipients. The effects are antigen-specific. The experiments described in this report are a comparison of the relationship of the route of administration of various transfer factors to the magnitude of the delayed hypersensitivity responses (footpad swelling) to the corresponding antigen in the recipients. Three doses of each of four affinity-purified transfer factor preparations were studied. There were no significant differences in the footpad responses by recipients of either oral or subcutaneous transfer factor. These results support proposals for oral administration of transfer factors in clinical trials.

Monoclonal antibodies defining functional sites on the toxin superantigen staphylococcal enterotoxin B.

Four monoclonal antibodies (mAbs) were produced binding to four nonoverlapping epitopes on the superantigen staphylococcal enterotoxin B (SEB). The mAbs were tested for their ability to detect SEB bound to major histocompatibility complex (MHC) class II, to inhibit SEB binding to MHC class II, to inhibit SEB stimulation of T cell hybridomas, to bind to various nonfunctional mutants of SEB, and to capture and present SEB and its mutants to T cells in the absence of MHC class II. We concluded that two mAbs, B344 and B327, bound to epitopes not required for superantigen function, one mAb, 2B33, blocked an MHC interaction site on SEB, and the fourth mAb, B87, blocked the T cell recognition site on SEB. Moreover, two mAbs (B344 and 2B33) were capable of presenting SEB, although much less efficiently than APC, to CD4- but not CD4+ T cell hybridomas. The results confirm the functional domains on SEB originally defined by mutation and show that MHC class II is not always an essential component of the superantigen ligand.

Obsessive compulsive disorder.

Clinical features of obsessive compulsive disorder (OCD) were found to be similar in various cultures. However, there was no report about phenomenology of this disorder from Moslem or Arab cultures. This study is a review of 45 cases who presented as a psychiatric clinic at a general university hospital in Saudi Arabia. The findings were found to be similar to those reported in Western studies with regard to age of onset, level of functioning, type of onset, course, and co-morbidity. Religious obsessions and compulsions were found to be the most common clinical features. Findings are explained in cultural terms.

The seasonality of mania: Preliminary findings.

The relationship between climatic variables and monthly manic admissions to a teaching hospital in Riyadh, Saudi Arabia was examined over a six-year period. The results indicate a weak but significant correlation between monthly admission rates of mania and day length, humidity, air pressure, and temperature. The correlation did not seem, however, to be strong enough to indicate a seasonal trend. Also, current month climatic variablesare better correlated than previous months. The prevalence of sunshine throughout the year may explain the low correlation with mania admissions unlike reports from other countries.