bluuebunny/crossref_metadata_2025 huggingface dataset:
Raised levels of tumour necrosis factor-alpha and neopterin, but not interferon-alpha, in serum of HIV-1-infected patients from Ethiopia
Authors: AYEHUNIE S , SONNERBORG A , YEMANE-BERHAN T , ZEWDIE D W , BRITTON S , STRANNEGARD O
Published: July 2010

Serum levels of tumour necrosis factor-alpha (TNF-α). neopterin and interferon-alpha (IFN-α) were determined by immunoradiometric assays in 60 HIV-1+and 20 HIV-1− subjects from Ethiopia. Swedish samples were used as reference material. The Ethiopian HIV-1+ subjects were found to have significantly increased TNF-α and neopterin, but not IFN-α levels. Increased levels of TNF-α and neopterin were frequently found in Ethiopian asymptomatic subjects (37% and 41%). and the concentration increased in patients with AIDS (83% and 90% respectively). The levels of the two substances and the proportion of patients with higher TNF-α values were lower in the corresponding Swedish subjects. The proportion of sera with raised levels of I FN-α was very low (asymptomatic 4%, and AIDS 7%) in Ethiopian subjects. These results suggest a very early increase in the TNF-α production and activation of the cellular immune response, and a low level of IFN-α synthesis in the natural course of HIV infection in Ethiopia. The aberrations may contribute to a rapid progress of immunodeficiency and cachexia often seen in Ethiopian patients.
Relationship ofin vitrophagocytosis of serotype 14Streptococcus pneumoniaeto specific class and IgG subclass antibody levels in healthy adults
Authors: LORTAN J E , KANIUK A StC , MONTEIL M A
Published: July 2010

The role of specific IgG2 antibody in the protection against serious infection with Streptococcus pneumoniae is unclear. We therefore decided to investigate the relationship between serum antibody levels and opsonization and phagocytosis of this microorganism. We have measured serum IgM, IgA and IgG subclass antibody specific for pneumococcal capsular polysaccharide and in vitro phagocytosis of serotype 14 pneumococcus by polymorphs, in healthy adults before and after immunization with Pneumovax II. IgM and IgG2 were the predominant anti-pneumococcal antibodies seen, IgA and IgGl being present at low titre. No significant relationship of phagocytosis with specific IgM and IgA antibodies was found. However, both specific IgG 1 and IgG2 antibodies in post-immunization sera correlated significantly with phagocytosis of the pneumococcus in the presence of complement (r= 0.57, P= 0.029 and r= 0.59, P= 0.022 respectively). After heat-inactivation, the remaining opsonic activity of sera correlated only with levels of specific IgG2 antibody (r= 0.61, P = 0.0006). Whereas phagocytosis supported by specific IgG1 and IgG2 antibody to serotype 14 pneumococcus after immunization is mediated by complement activation, IgG2-specific antibody in high titre may also be able to function by complement-independent interaction with Fcγ receptors on polymorphs.
Quantification of antibody-secreting lymphocytes that react with Pf155/RESA fromPlasmodium falciparum: an ELISPOT assay for field studies
Authors: FIEVET N , CHOUGNET C , DUBOIS B , DELORON P
Published: July 2010

We have adapted the enzyme-linked immunospot assay (ELISPOT) to enumerate the cells from Plasmodium falciparum-primed donors that produce IgG in vitro in response to malaria antigens. In vitro activation of cell cultures with two synthetic peptides (EENVEHDA)4, and (LGRSGGDIIKMQTL) corresponding to immunodominant T cell epitopes of the ring-infected erythrocyte surface antigen (Pf 155/RESA) gave specific antibody-secreting cells (ASC) in five and six of the 15 P. falciparum-primed donors from Cameroon. Antibodies produced after a stimulation by synthetic peptides reacted also with total parasite proteins. However, crude P. falciparum antigen did not trigger a higher number of cells than did synthetic peptides. The absence of significant relation between the presence of sera antibodies and in vitro ASC against the same peptide suggests that the kinetics of circulating primed lymphocytes and antibodies are different. We evaluated 0–04–0–29% of peripheral blood B cells to be the frequency of memory cells specific to a single Pfl 55/RESA epitope in these donors. This study suggests that the ELISPOT assay should permit the analysis of B cell responses to malarial antigens at the single-cell level and its applicability to epidemiological field studies. This assay should be well suited to the identification of T helper epitopes capable of inducing the production of antibodies by human B cells, and will constitute an important tool for the selection of immunogens to be included in a subunit vaccine.
Specific increases in urinary excretion of anti-DNA antibodies in lupus mice induced by lysozyme administration: further evidence for DNA-anti-DNA immune complexes in the pathogenesis of nephritis
Authors: YAMAMOTO T , NAGASE M , HISHIDA A , HONDA N
Published: July 2010

We previously reported that lysozyme electrostatically inhibits the fibronectin-mediated DNA binding to the glomerular basement membrane (GBM) and reduces in situ DNA-anti-DNA complex formation in the GBM in NZB/W F1 mice [I]. In this study, we further noticed significant increases in urinary excretion of anti-DNA antibodies and immune complexes (IC) in lysozyme-treated NZB/W F1 mice. Their clearance ratios of IgG anti-DNA antibody to whole IgG were markedly high compared with those of saline-treated animals. A large number of IgG and C3 positive granules were observed in the tubular cells of NZB/W F1 mice treated with lysozyme. On the contrary, nil or only small amounts of anti-DNA antibodies were detected in the urine of NZB/W F1 mice without lysozyme administration despite a large amount of proteinuria, suggesting entrapment of the antibodies in lupus glomeruli. Lysozyme neither inhibited the binding of anti-DNA antibodies to DNA or heparan sulphate nor did it displace anti-DNA antibodies and IC from the kidney homogenates of lupus mice. It thus appears that the inhibition of DNA binding to the GBM due to lysozyme reduced the entrapment of anti-DNA antibodies in the GBM, resulting in urinary excretion of the antibodies.
Effect of chlorpromazine on kinetics of injected monoclonal antibody in MoAb-induced glomerular injury
Authors: TAKASHIMA N , KAWACHI H , OITE T , NISHI S , ARAKAWA M , SHIMIZU F
Published: July 2010

The effect of chlorpromazine, one of several calmodulin antagonists that inhibit cytoskeletal movement, on the local kinetics of injected proteinuria-inducing MoAb 5–1–6 was examined to test the hypothesis that proteinuria is inhibited if the antigen recognized by MoAb 5–1–6 or injected MoAb remains on the surface of epithelial foot processes. MoAb 5–1–6 was injected into both chlorpromazine-treated (5 mg/100 g body weight) and untreated rats. As a positive control for the chlorpromazine treatment, anti-Fx 1A serum was also injected into other chlorpromazine-treated and untreated rats. Chlorpromazine inhibited neither the change in localization of injected MoAb 5–1–6 nor proteinuria, although it showed an inhibitory effect on redistribution of immune complex and the fixation of complement in passive Heymann glomerulonephritis induced by injection of anti-Fx 1A serum. We conclude that the kinetics of bound MoAb 5–1–6 are regulated by a system different from that operating in passive Heymann glomerulonephritis.
Asp57-negative HLA DQβ chain and DQA1*0501 allele are essential for the onset of DQw2-positive and DQw2-negative coeliac disease
Authors: MANTOVANI V , CORAZZA G R , BRAGLIANI M , FRISONI M , ZANIBONI M G , GASBARRINI G
Published: July 2010

The genetic predisposition to coeliac disease is associated with the HLA DQw2 allele. Coeliac patients lacking the DQw2 allele are very rare and always exhibit the DR4-DQw3 haplotype. We performed oligotyping of polymerase chain reaction (PCR)-amplified DQA1 and DQB1 genes in six DQw2-negative and 30 DQw2-positive coeliac patients. The DQB analysis showed that all six DQw2-negative patients possessed the DQB 1*0302 allele. The other DQB alleles found in five of these patients were DQB1*0501, DQB1*0604 and DQB1*0302. The DQβ chains encoded from all these alleles have the replacement of aspartic acid residue at position 57 (Asp57), as well as the DQB 1 *0201 allele which was found in all 30 DQw2-positive coeliac patients. The DQw2-negative proband who lacked the homozygous Asp57 replacement exhibited the DQA1*0501 allele in the DQA1 gene. The DQA1 *0501 allele was also found in 27 of the 30 DQw2-positive coeliac patients. Among this group of coeliacs, the four cases lacking the DQA1*0501 allele exhibited the homozygous Asp57 replacement in the DQβ chain. Our results indicate that Asp57-negative DQβ alleles are involved in both DQw2-positive and -negative coeliac patients. Moreover, when the Asp57-negative DQβ chain is encoded from only one of the two DQB1 genes the DQA1*0501 allele is always present.
Activation of the immune system of cancer patients by continuous i.v. recombinant IL-2 (rIL-2) therapy is dependent on dose and schedule of rIL-2
Authors: GRATAMA J W , BRUIN R J , LAMERS C H J , OOSTEROM R , BRAAKMAN E , STOTER G , BOLHUIS R L H
Published: July 2010

The effect of dose and schedule of continuous i.v. rIL-2 infusions on leucocyte subset counts, activation status of CD56+CD3- natural killer (NK) and CD3+ T lymphocytes, and cytolytic activities of peripheral blood mononuclear cells (PBMC) was studied. A single 4-day course of rIL-2 in escalating doses (0.9-11.5 × 106 U/m2 per day) was given to 18 patients with various types of metastatic cancer. The serum IL-2 concentration during rIL-2 Iherapy ranged between 23 and 64 U/ ml and was proportional to ihe administered rIL-2 dose, as was the rebound lympbocytosis following therapy. Before therapy, the CD56+CD3- NK cells expressed low levels of the p75 chain of the IL-2 receptor (IL-2R) and virtually no IL-2R(p55). Most CD3+ T cells were IL-2R(p55,p75). Between 2 and 4 days following therapy, i.e. at the time of lymphocytosis, the percentage of CD56+, CD3 NK cells among tbe lymphocytes had increased proportional to tbe administered rIL-2dose. The levels of IL-2R(p75) expression by the CD56+, CD3 NK cells had increased. The percentages of CD3+ T cells expressing IL-2R(p55). H LA-DR and CD45RO had increased proportional to the administered rIL-2 dose. The level of lymphokine- activated killer (LAK) activity against Daudi cells was also positively correlated with rIL-2 dose. Subsequently, seven patients received 4-weekly cycles of rIL-2 (2.9-4.4 × 106 U/m2 per day) during 4 consecutive weeks. This schedule led to marked increments in lymphocyte and eosinopbil counts, and to increased cytolytic activities compared with pretreatment. We conclude that CD56+, CD3- NK and CD3+ T cells are activated differentially by continuous i.v. rIL-2 proportional to dose and duration of treatment.
Serum and salivary responses to oral tetravalent reassortant rotavirus vaccine in newborns
Authors: FRIEDMAN M G , SEGAL B , ZEDAKA R , SAROV B , MARGALITH M , BISHOP R , DAGAN R
Published: July 2010

Serum and salivary responses of 95 infants to either a standard (4 ± 104 plaque-forming units (PFU), 47 neonates) or a high dose (4 ± 105 PFU, 48 neonates) of tetravalent reassortant rhesus rotavirus vaccine (administered at 2 days and at 6 weeks of age) were evaluated in a double-blind clinical trial. Serum and salivary IgA antibodies to the rotavirus group A common antigen were determined by ELISA and radioimmunoassay (RIA). Serum neutralizing antibodies to rhesus rotavirus were determined by fluoreseent foeus reduction assay. No significant differences in responses to the high versus standard dose were noted in serum or saliva. Response was influenced by cord blood antibodies. All infants who were cord blood-negative for rhesus rotavirus neutralizing antibodies (nine who received the standard dose and 20 who received the higher dose) had serum responses, compared with 42-70% of those who were cord blood-positive. The serum response rate recorded for babies with cord blood neutralizing titres > 1000 was 44%. Infants being bottle fed had a higher serum response rate than did babies being breast fed exclusively. If serum and salivary responses were combined, the response rate reached 80% for bottle fed infants. Thus, determination of serum responses alone underestimates vaccine ‘take’ in infants, and more so in highly endetnie areas thati in areas subject only to sporadic outbreaks. However, determination of salivary responses in newborn breastfed infants may be inaccurate, due to possible persistence of antibodies derived frotn colostrum or breast milk.
Lymphocyte changes associated with prolongation of cardiac allograft survival in adult mice using anti-CD4 monoclonal antibody
Authors: PEARSON T C , BUSHELL A R , DARBY C R , WEST L J , MORRIS P J , WOOD K J
Published: July 2010

This study investigated the effect of anti-CD4 MoAb treatment on lymphocyte phenotype and function and correlated these changes with the prolongation of cardiac allograft survival in adult mice. Indefinite survival of hcterotopic cardiac aliografts was obtained in several fully allogencie strain combinations when two doses of the anti-CD4 MoAb, YTS 191.1, were given at the time of transplantation, A dose response analysis in the C57BL/10 to C3H/He strain combination showed that very low doses of YTS 191.1 (25 μg/dose) were able to induce prolonged allograft survival when administered perioperatively. At the time of transplantation the immunosuppression induced by administration of the anti-CD4 MoAb is not antigen-specific, as heart grafts from different donor strains, mismatched for both major and minor histoeompatibility antigens, showed prolonged survival in treated recipients. Immunocompetence was restored by 6 weeks after MoAb treatment, as recipients regained the ability to reject a cardiac allograft transplanted at this time point. However, while recovery of immunocompetence could be demonstrated in civo, leucocytes isolated from the peripheral lymphoid organs of treated mice continued to be hyporesponsive in mixed leucocyte culture (MLC), Phenotypic analysis of the peripheral lymphoid tissues showed that C3H/He recipients treated with 25 μg/dose of YTS 191.1 had a marked, but not complete, elimination of the CD4+ subset al the time of transplantation, which was gradually restored to 50% of normal by 6 weeks after treatment. Thus, complete elimination of the CD4+ subset was not required to achieve indefinite allograft survival, and immunocompetence, as assessed in vivo, returned even when the CD4+ subset was present at half the normal level. Low doses of anti-CD4 MoAb (25 μg) had no effect on the expression of the CD4 molecule by thymocytes, and yet thymocytes were hyporesponsive to alloantigen in vitro. At higher doses of YTS 191.1, immature CD4+ 8+ thymoeytcs were selectively depicted. These results suggest that anti-CD4 MoAb therapy may modulate the intrathymic T cell selection process. These studies provide further insight into the mechanism of action of low dose, depleting anti-CD4 MoAb therapy in allograft rejection, and form a basis from which rational modifications to therapeutic protocols in transplantation models can be made.
Therapeutic effect of granulocyte colony-stimulating factor and cephem antibiotics against experimental infections in neutropenic mice induced by cyclophosphamide
Authors: WAKIYAMA H , TSURU S , HATA N , SHINOMIYA M , SHINOMIYA N , NORITAKE M , UMEZAWA Y , ROKUTANDA M
Published: July 2010

Effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) against severe infections in cyclophosphamide (CY)-induced neutropenic mice were investigated by its single use or by its combination with cephem antibiotics. Treatment with rhG-CSF increased the number of peripheral blood leucocytes and strikingly shortened the duration of the neutropenic state. Most of the regained population in the peripheral blood leucocytes were neutrophils. Functions of neutrophils, such as phagocytic activity, superoxide release, and expression of Mac-1 molecules, were remarkably enhanced by administration of rhG-CSF. When rhG-CSF was administered to CY-induced neulropenic mice before infection, protective effects against various kinds of bacteria were remarkable. On the other hand, such remarkable effects were not observed when rhG-CSF was administered after infections. However, even in the case of post-infectious treatment, a combination therapy of rhG-CSF with cephem antibiotics, particularly with Cefodizime (CDZM), showed a significant improvement in the survival rate and a decrease in the number of viable bacteria in the liver. These results suggest that a combination therapy of rhG-CSF with cephem antibiotics, especially with CDZM, is an efficient regime against severe infections in patients with ncutropenia induced by a heavy anti-tumour chemotherapy.
Isotype-specific cross-linking of select human FcγR isoforms triggers release of IL-6
Authors: DUITS A J , AARDEN L A , ERNST L K , CAPEL P J A , van de WINKEL J G J
Published: July 2010

Anti-CD3 MoAbs arc widely used in T cell activation studies, and are effective in immunosuppressive therapy. We used a panel of mouse (m) anti-CD3 switch variant MoAbs of five different isotypes to study IL-6 release from accessory cells. Incubation of human (h) mononuelear cells with anti-CD3 MoAbs resulted in increased IL-6 levels with MoAbs of mIgG 1 and mIgG2a isotypes. with no effect of mlgG2b or mIgA. This suggested involvement of IgG Fc receptors (FcγR) in triggering IL-6 production. To evaluate the role of different FcγR molecules individually we used a panel of hFcγR-transfeeted mouse fibroblasts, and Jurkat T cells as a model. IL-6 secretion by CD32 transfectants expressing the hFcγRIIa high-responder (HR) allelic form was triggered by mIgG1 anti-CD.3 MoAb. with no effect of four other isotypes. None of the anti-CD3 MoAbs induced IL-6 secretion by CD32 transfectants expressing either a variant of this receptor, containing only a single intracellular amino acid (CT-), the hFcγRIIa low-responder (LR) allelic form, or hFcγRIIbl. hFcγRI (CD64) transfectants exhibited IL-6 production after incubation with mlgG2a anti-CD3 MoAb, and to a lesser extent with mlgG2b. and mlgG I MoAb. Indirect involvement of T cells in triggering IL-6 secretion could be excluded by experiments in which transfectants were cultured with immobilized anti-CD3 MoAb. These data indicate that cross-linking of cither hFcγRI, or hFcγRIIaHR by appropriate anli-CD3 MoAbs triggers IL-6 production of accessory cells, and not T cells. This may also take place in vivo during immunosuppressive therapy with anti-CD3 MoAbs, and related antibody-mediated immune responses.
The anti-lipid A antibody HA-IA binds to rough Gram-negative bacteria, fixes complement and facilitates binding to erythrocyte CRl (CD35)
Authors: TONOLI M , DAVIES K A , NORSWORTHY P J , COHEN J , WALPORT M J
Published: July 2010

MoAbs to bacterial cell wall Iipopolysaccharidc arc currently under evaluation for the treatment of Gram-negative sepsis. The mode of action of these reagents remains poorly understood. In this study we examined the ability of radiolabelled HA-IA (an IgM anti-lipid A MoAb) to bind in vitro to Salmonella minnesota (Re 595), Escherichia coli, and Streptococcus pyogenes. HA-1A was able to bind specifically to the “rough” mutant Stilm. minnesota. but not to a ‘smooth’E. coli, or Strep, pvogenes. Binding to Salm. minnesota led to complement fixation which resulted in bacterial adherence to erythrocyte CR1, suggesting a possible mechanism whereby the antibody might enhance clearance of bacteria by facilitating delivery to the fixed mononuclear phagocytic system. We were not able to demonstrate the formation of immune complexes between free lipopolysaccharide and HA-IA in the presence of serum, nor the enhancement of complement-mediated binding of HA-1 A:Salm. minnesota immune complexes to erythrocytes by antibiotic treatment. Binding of HA-IA to small bacterial fragments was, however, demonstrable after in vitro treatment with a β-lactam antibiotic, which disrupts the bacterial cell wall, but not with gentamicin, an aminoglycoside antibiotic which blocks protein synthesis.
Synovial fluid antigen-presenting cell function in rheumatoid arthritis
Authors: VINER N J , GASTON J S H , BACON P A
Published: July 2010

We have previously demonstrated enhanced synovial fluid (SF) antigen-presenting cell (APC) function in inflammatory arthritis patients selected on the basis of marked SF mononuclcar cell (MNC) responsiveness to reactive arthritis-associated bacteria (Clin Exp Immunol 1990; 79:189-94). In this study we have assessed whether similarly enhanced synovial APC function is present in other inflammatory arthritis patients by using two assay systems to study 18 rheumatoid arthritis patients whose MNC responsiveness had not been determined in advance. We demonstrate that rheumatoid SF APC are much more potent than peripheral blood (PB) APC in stimulating the responses of autologous PB T cells to a range of recall antigens. In addition, SF APC are shown to be efficient stimulators of the antigen-specific responses of MHC-compatible, cloned T cells. Enhanced synovial APC function is thus likely to be a general feature of inflammatory arthritis and may play an important role in its pathogenesis.
γσ Lymphocytes in endocrine autoimmunity: evidence of expansion in Graves' disease but not in type 1 diabetes
Authors: ROURA-MIR I C , ALCALDE L , VARGAS F , TOLOSA E , OBIOLS G , FOZ M , JARAQUEMADA D , PUJOL-BORRELL R
Published: July 2010

Endocrine autoimmune disorders are mediated by T cell-dependent responses to organ-specific antigens, but the mechanisms initiating the process remain unknown. Lymphocytes whieh use the γδ heterodimer as T ceii receptor (TCR) for antigen constitute a distinct subset of T cells whose function remains elusive. In order to investigate their possible involvement in endocrine auloimmunity we have determined the proportion of γδ T cells in the peripheral biood of 23 patients with type 1 (insulin-dependent) diabetes mellitus (type-1 DM) and 30 patients with autoimmune thyrotoxicosis (Graves’ disease). T lymphocyte TCR expression was assessed by fluorescence-activated flow eytometry on peripheral blood mononuclear cells using MoAbs UCHTI (CD3), TCR 51 (γδ TCR), WT31 and βF1 (αβ TCR) and both the percentage of T cells expressing γδ and the ratio γδ/αβ were calculated. In the diabetie patients γδ cells were not significantly different from (he control group (7·7 ± 54%versus 8·0 ± 5·5%) of T eells, P NS). There was no relation between the proportion of γδ lymphoeytes and the presence ol’ islet cell antibodies (ICA) in the sera. The Graves’ patients showed a tendency towards a higher proportion of γδ T lymphocytes than the controls (γδ/αβ ratios: 0·095 ± 0·047 versus 0·063 ± 0·022, P= 0·03). In 14 Graves’ patients the number of γδ were measured in paired samples of peripheral and inlrathyroidal lymphocytes, demonstrating an expansion of γδ within the thyroid glands (0·21 ± 0·3 versus 0·095 ± 0·047, P= 0·032). Immunohistochemical studies showed that γδ celts were scattered among the predominant αβ lymphoeytes infiltrating the thyroid gland and that they aeeount for 10% of intraepitheliai lymphocytes. No relation was found between the increase of γδ lymphocytes and any clinieai features.
Trafficking of inflammatory macrophages from the kidney to draining lymph nodes during experimental glomerulonephritis
Authors: LAN H Y , NIKOLIC-PATERSON D J , ATKINS R C
Published: July 2010

Macrophage accumulation within the glomerulus and renal interstitium is a prominent feature of most forms of glomerulonephritis, but the fate of these inflammatory cells is unknown. Macrophage trafficking to the draining kidney lymph nodes (KLN) was assessed in a detailed kinetic analysis of accelerated antiglomerular basement membrane (GBM) disease in the rat. Leucocytes draining to KLN via lymphatic vessels were identified within the marginal sinus by MoAb labelling of tissue sections. In anti-GBM disease, there was a significant increase in the weight of the KLN due to both lymphoproliferation within the nodes and increased lymphatic drainage from the inflamed kidney, as evidenced by prominent dilation of the marginal sinus and increased numbers of cells within the sinus. In non-inflamed lymph nodes, few ED1+ macrophages were present within the marginal sinus (3·0±0·6/100 nucleated cells). However, in anti-GBM disease, macrophages became the major cell type within the dilated marginal sinus of the KLN, as shown by labelling with ED1, ED2 and ED3 MoAbs, peaking at 74±2·6 ED1+ cells/100 nucleated cells at day 14. These changes were not simply due to systemic antigen administration, since in the axillary lymph node (ALN) there was no obvious dilation of the marginal sinus and macrophages accounted for a maximum of only 15±4·6 ED1+ cells/100 nucleated cells. In conclusion, this study provides indirect evidence that there is significant trafficking of the renal macrophage infiltrate to the KLN during experimental glomerulonephritis. This may be a mechanism whereby nephritogenic antigens, released as a consequence of the local inflammatory response, may be presented to T and B lymphocytes within lymph nodes, resulting in the amplification of the immune response in glomerulonephritis.
Quantitative study of mesangial injury with proteinuria induced by monoclonal antibody 1-22-3
Authors: KAWACHI H , OITE T , SHIMIZU F
Published: July 2010

Murine MoAb 1-22-3 has already been reported to bind to the mesangial cell surface and to cause transient proteinuria and mesangial morphological changes characterized by mesangiolysis. subsequent mesangial cell proliferation and mesangial matrix increase by a single i.v. injection. In this study, MoAb-induced glomerulopathy was quantitatively analysed. No correlation between the severity of mesangial morphological changes and the degree of proteinuria was detected (r= 0·190). The minimum dose injected to induce abnormal proteinuria was 25 μg. This dose corresponded to 1·79 μg/2 kidneys 30 min after MoAb injection. The highest average level of proteinuria was observed in rats injected with 500 μg of MoAb, and less proteinuria was observed in rats injected with 10·0, 5·0 and 2·0mg. Although the amounts of kidney-fixing MoAb and the subsequent deposition of rat C3 in the high-dose-injected group were larger than in the 500 ng injected group, the numbers of infiltrating inflammatory cells were the same in both groups. No correlations between the degrees of such mediators and proteinuria were observed.
Mucosal macrophage subsets of the gut in HIV: decrease in antigen-presenting cell phenotype
Authors: LIM S G , CONDEZ A , POULTER L W
Published: July 2010

The effect of HIV infection on intestinal lamina propria macrophage subsets was investigated in 41 patients at various stages of HIV infection (asymptomatic HIV infection, n= 17; AIDS, n= 24). Duodenal biopsies taken from HIV patients at endoscopy were snap frozen and cryostat sections cut for immunohistochemical staining. MoAbs CD68 (EBM11. pan-macrophage marker). RFD1 (antigen-presenting cells) and RFD7 (mature phagocytic macrophages) were used to identify cell subsets using indirect immunoperoxidase or alkaline phosphatase. Double immunofluoresccncc using MoAbs to HIV proteins (p24, p17 and gp120) and RFD1 were used to identify HIV-infected antigen-presenting cells. Double immunofluoresccncc was also used to identify macrophages that expressed both RFD1 and RFD7 (‘suppressor’ macrophages). Intensity of HLA-DR expression in lamina propria cells was investigated using a MoAb to HLA-DR directly conjugated to glucose oxidase. The results show that there was no difference in overall density of macrophages, but there was a significant decrease in dendritic cells (RFD1+) in all clinical stages of HIV. There was no difference in the density of RFD7+ macrophages, nor was there a difference in intensity of HLA-DR expression in lamina propria cells. Only four HIV-infected cells were positively identified in the 41 patients. This result suggests that the antigen-presenting arm of mucosal immune defences may be seriously compromised in HIV infection, and represents a further insult to mucosal immunity already impaired as a result of loss of CD4+ T lymphocytes. This may contribute to development of opportunist infection in the gut.
In vitroimmunotoxicity of ±2′-deoxy-3′-thiacytidine, a new and-HIV agent
Authors: LISIGNOLI G , MONACO M C G , DEGRASSI A , TONEGUZZI S , RICCHI E , COSTIGLIOLA P , FACCHINI A
Published: July 2010

The present study compares the in vitro effect of (±)-2′-deoxy-3′-lhiacylidine (BCH 189) a new synthetic anti-HIV-l dideoxynucleosidc. with 3′-azido-3′-deoxythymidinc (AZT) on the immune function of lymphocytes from 10 normal and 12 HIV-1+ patients (CDC II and III). The effect of different doses of BCH 189 and AZT was analysed in vitro on: (i) T cell proliferation after stimulation with concanavalin A (Con A) or anti-CD3 MoAb: (ii) B cell proliferation and immunoglobulin production after stimulation with pokeweed mitogen (PWM); (iii) cytokine production (IL-2. IL-6. GM-CSF, tumour necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ)) from lymphocytes stimulated with anti-CD3 MoAb or phylohucmagglutinin (PHA). BCH 189 inhibited the proliferation of B and T lymphocytes from normal and HIV’ subjects less than AZT: even if lymphocytes from HIV+ (CDC HI) subjects produced higher levels of IL-6 and TNF-α, neither BCH 189 nor AZT molecule interfered with cytokine release. Immunoglobulin production from B lymphocytes was inhibited only by a high concentration (50 μm) of BCH 189 or AZT. These results show that BCH 189 affects lymphocyte proliferation in vitro less then AZT, and support its use in clinical trials in HIV-infected patients.
Elevated levels of antibodies against sulphatide are present in all chronic chagasic and dilated cardiomyopathy sera
Authors: AVILA J L , ROJAS M , CARRASCO H
Published: July 2010

A natural anti-sulphatide antibody was found to be present in the serum of every normal individual studied. The reactivity of the antibody was assessed by its interaction with galactosylceramide-P-sulphate. Antigen-antibody binding was strongly blocked by 1 mm heparin, dextran sulphate and chondroitin sulphate A, and by 5 mM chondroitin sulphate B. Antibodies avidly absorb to rabbit erythrocytes, but discretely to rat erythrocytes, suggesting that they are different from galactocere-broside antibodies. Elevated levels of sulphatide antibodies were present in all of 102 chronic Trypanosoma cruzi-infected patients studied, but not in other patients having cutaneous or visceral leishmaniasis, T. rangeli infection or several other protozoal, helminthic or mycotic infections. Interestingly, 100% of 40 dilated cardiomyopathy patients also have elevated levels of sulphatide antibodies. As T. cruzi is rich in galactocerebroside sulphate, it is proposed that in chagasic patients this glycolipid could act as an immunogen, inducing elevated titres of sulphatide antibodies, which could be important in the pathogenesis of cardiac or peripheral nerve symptoms.
Failure of FK 506 to suppress the T cell-mediated immunity of mice toListeria monocytogenes
Authors: WAGNER J A , KRETSCHMAR M , NICHTERLEIN T , HOF H , QUADE B
Published: July 2010

Listeria monocytogenes belongs to the group of intracellular bacteria, which means that they reside and multiply within host cells. The protective immunity against such an infection is mediated by cellular immune mechanisms. Whereas the CD8+ T cell population plays a major role therein, the CD4+ T cells are held to be of minor importance in this defence system. Consequently, one can understand that immune suppression with FK 506 which acts primarily on this latter T cell subset, docs not interfere with protective immunity of mice infected with L. monocytogenes. We have demonstrated that the drug blocks neither curing of primary infection, nor formation of granulomas, nor induction of cell populations capable of mediating adoptive transfer of immunity, nor expression of pre-existing immunity.
Early IL-2/sIL-2R surge following surgery leads to temporary immune refractoriness
Authors: LAHAT N , SHTILLER R , ZLOTNICK A Y , MERIN G
Published: July 2010

High serum level of immunoreactive but not biologically active IL-2 was detected 1 day after surgery in patients undergoing major operation (abdominal, open-heart), in proportion to the tissue injury caused by surgical trauma. IL-2 values were highest in those patients who underwent open-heart surgery and received blood transfusions. In all patients they declined in the third and fourth postoperative days. Elevated serum levels of soluble IL-2 receptors (sIL-2R) were already present 1 day after operation, and peaked in the third and fifth post-opperative days after mitogen triggering. Blood lymphocytes derived from operated patients secreted reduced amounts of bolh IL-2 and sIL-2R compared with control lymphocyies. The extent and duration of this reduction were also proportional to the tissue trauma and were affected by blood transfusions. Based on these data we suggest that early post-operative systemic immunological activation (appearance of IL-2 in the serum) is followed by elevation of sIL-2R. which then interferes with IL-2-dependent immunity. Blood lymphocytes are probably not involved in the post-operative immunological activation. The trigger for and the site of IL-2 /s IL-2R synthesis are not yet clear.
Ultrastructural studies bearing on the mechanism of UVB-impaired induction of contact hypersensitivity to DNCB in man
Authors: MOMMAAS A M , MULDER A A , VERMEER M , BOOM B W , TSENG C , TAYLOR J R , STREILEIN J W
Published: July 2010

In both murine and human experimental systems, acute, low dose exposure of skin to ultraviolet B light (UVB) impairs the induction of allergic contact dermatitis (ACD) by haptens such as dinitrochlorobenzene (DNCB) in a significant proportion of individuals. By light microscopy, epidermal Langerhans cells (LC) have been reported to be depleted by UVB exposure as well as by epicutaneous hapten application, implying that LC may be the locus of action of the effeeis of both UVB and DNCB. However, light microscopy can not readily distinguish cell density changes secondary to LC necrosis from changes resulting from down-modulation of expression of LC surface molecules. Using a highly sensitive immunogold electron microscopic approach, we have evaluated the differential effects of UVB and/or DNCB on human epidermal LC. The results reveal that DNCB alone eaused significant up-regulation of cell surface HLA elass II expression on a very small number of LC. the major fraction of LC expressing normal levels of HLA class II. Furthermore, DNCB alone caused a modest reduction in the density of LC at the treated sites without evidence of cell necrosis. Treatment with UVB alone or UVB exposure followed by DNCB resulted in a reduction in the density of LC. with widespread evidence of LC necrosis. However, the few remaining intact LC were all intensely HLA class II-positive after UVB exposure followed by DNCB. whereas treatment with UVB alone did not result in changes in LC HLA class II expression. The findings that after DNCB painting only a small proportion of the LC were strongly HLA class II-positive. but after UVB exposure followed by DNCB all intact LC displayed significant up-regulation of cell surface HLA class II expression, imply that UVB exposure inhibits the migration of epidermal LC. This is consistent with the view that DNCB fails to induce ACD when hapten is painted on UVB-exposed skin because insufficient LC are available to initiate T cell activation in the draining lymph node.
Concomitant enhancement of the response to Mls-la antigens and the induction of post-thymectomy autoimmune gastritis in BALB/c mice
Authors: MURAKAMI K , HOSONO M , MARUYAMA H , MORI Y , NISHIO A , FUKUMOTO M , WATANABE Y , INABA M , KURIBAYASHI K , SAKA M , MASUDA T
Published: July 2010

We examined the role of Mis antigens in the induction of autoimmune gastritis (AIG) in BALB/c and DBA/2 mice subjected to thymeclomy. The prevalence of AIG in Mls-lb mice which underwent thymectomy on day 3 after birth (3d-Tx) was 78% (mean), while in Mis-1a DBA/2 mice it was < 6%. Whereas AlG-negative 3d-Tx DBA/2 mice produced 2-mercaptoethanol (2-MF.)–sensitive anti-parietal cell autoantibody. AIG-positive BALB/c mice made 2–ME-resistant anti-parietal cell autoantibody. In addition, the prevalence of AIG in 3d-Tx BALB/c mice which were rendered tolerant to M Is-1a antigens by injection of bone marrow cells from (BALB/c ± DBA/2)F1 mice within 24 h after birth was decreased compared with the non-tolerant control mice: the prevalence being 80% in the controls and 30% in the tolerant animals. Thus, the activation of helper T cells, including T cells responding to Mls-1a antigens and including immunoglobulin class switch, appeared to be closely associated with the induction of AIG. Flowcytometric analysis confirmed that C'D4- Vβ6 T cells had increased in the regional lymph nodes of the stomach in AIG mice. However, an increase in the number of Vβll T cells, which are known to increase in 3d-Tx mice, occurred in the CD8, but not in the CD4 T cell population. Injection of MoAb to L3T4. but not Lyt2. Vβ6-or Vβ8–TCR, into 3d-Tx BALB/c and syngeneic nude mice which had received spleen cells of 3d-Tx BALB/c mice bearing AIG completely abrogated the development of AIG. despite there being remarkable decreases in T cells expressing relevant markers to the injected antibodies in all the mice. These findings suggest that the increase of V bT6+L3T4+ T cells in AIG mice was concomitant with the activation of A IG-inducing Vβ6- L3T4+ T cells.
An acute model for IgA-mediated glomerular inflammation in rats induced by monoclonal polymeric rat IgA antibodies
Authors: STAD R K , BRUIJN J A , van GIJLSWIJK-JANSSEN D J , van ES L A , DAHA M R
Published: July 2010

An acute model Tor IgA-mediated glomerular inflammation in rats was induced by the in situ deposition of IgA directly into the glomerular mesangium. F(ab')2 anti-Thyl MoAb was used to anchor an antigen. DNP (2,4-dinitrophenol), in the glomeruli of rats. Subsequent infusion of rat polymeric (p-) or monomeric (m-) IgA MoAb with specificity for DNP resulted in mesangial deposition of IgA in both groups of rats. However, acute proteinuria was observed only in p-IgA-treated rats and not in PBS- or m-IgA-treated rats. Immunofluorescence analysis revealed deposition of C3 in an identical pattern to that of IgA in the glomeruli of p-IgA-treated rats. No mesangial deposits of C4 or Clq were seen in these animals. Rats receiving m-IgA or PBS displayed no detectable C3, C4 or Clq deposition. The amount of proteinuria in p-IgA-treated rats was related to the amount of deposited C3. The presence of intraglomerular monocytes was only observed 2 days after p-IgA injection. By light microscopy, aneurysm formation, mesangial hypercellularity and matrix expansion were seen only in p-IgA-treated rats. However, by 37 days post-injection complete resolution of the lesions was observed. No histological renal changes were observed in PBS- or m-IgA-trcatcd rats. In conclusion, an acute form of IgA-mediated nephritis in rats was induced by p-lgA but not by m-IgA. This reproducible model provides a basis for further study into the mechanisms of IgA-mediated glomerular inflammation.
Comparison of potentiality to induce graft-versus-host reaction with small bowel, pancreas/spleen, and liver transplantation in the rat
Authors: KOBAYASHI E , KAMADA N , ENOSAWAJ S , TOYAMA N , DELRIVIERE L , GOTO S , KIM Y I L , MIYATA M
Published: July 2010

Although small bowel transplantation (SBT). or pancreas-spleen transplantation (PST) often lead to lethal graft-versus–host reaction (GVHR) in experimental animals, fatal GVHR is rare after clinical liver transplantation. This study describes a modified model of SBT and PST in the rat using cuff techniques applied to the renal artery and vein of the recipient. The ability of LEW (RTI1) or BN (RTln) lymphocytes accompanying intestinal, splenic, or hepatic grafts to induce lethal GVHR in (LEW BN) F1 hybrid recipients was compared. SBT and PST experiments showed that lethal GVHR always occurred in LEW-into-F1 combination, but was much less frequent in BN-into-F1 SBT. In mixed lymphocyte reaction (MLR), LEW mesenteric or splenic T cells showed significantly higher proliferative responses against BN stimulators than did BN mesenteric or splenic T cells against LEW. Adoptive cell transfer experiments using mesenleric or splenic cells also showed that LEW cells were higher responders than BN. In contrast with SBT and PST results, a lethal GVHR was not induced after liver or pancreas grafting alone in either parent-to-F1 combination. In MLR, hepatic T cells from either parent failed to elicit a proliferative response against allostimulators. These results indicate that the occurrence of lethal GVHR is dependent upon the reactivity of parental lymphocytes against allo-antigenicity of F1 hybrids and also upon the lymphoid tissue transplanted. The lack of alloreactivity of hepatic T cells accounts for the absence of lethal GVHR after liver grafting.
A soluble form of the human transferrin receptor is released by activated lymphocytesin vitro
Authors: WOITH W , NÜSSLEIN I , ANTONI C , DEJICA D I , WINKLER T H , HERRMANN M , PIRNER K , KALDEN J R , MANGER B
Published: July 2010

Soluble transferrin receptors (sTfR) were detected in culture supernatants of activated human peripheral blood mononuclcar cells (PBMC) using a sandwich ELISA technique with two non-cross-reacting TfR MoAbs. Mitogenic stimulation of lymphoid cells induced both up-regulation of TfR surface density and release of sTf R to the medium. Peak levels of sTfR in culture supernatants occurred at day 4 after activation, 1 day later than maximum expression of TfR in the plasma membrane. Production of sTfR was independent of proliferation, as demonstrated by measuring sTfR release by PBMC, which had becn irradiated with a dose of 20 Gy before activation. In addition to these in vitro experiments, we tested the sera of 85 patients with systemic lupus erythematosus (SLE). an autoimmune disease accompanied by in vivo activation of lymphocytes, for their sTfR levels. No correlation of these data was detectable to serum concentrations of the soluble α-chain of the IL-2 receptor, an unequivocal marker of lymphocyte activation. However, they correlated negatively to the haemoglobin content of the patients’ erythrocytes. indicating that erythroid progenitors are the predominant source of sTf R in SLE patients’ sera.
Subsets of CD8+, CD57+ cells in normal, healthy individuals: correlations with human cytomegalovirus (HCMV) carrier status, phenotypic and functional analyses
Authors: WANG E C Y , TAYLOR-WIEDEMAN J , PERERA P , FISHER J , BORYSIEWICZ L K
Published: July 2010

Two different subsets of CD8+, CD57+ cells have been defined, one expressing high levels (CD8high+(CD57+)), the other expressing low levels of surface CD8 (CD8low+(CD57+)). Increased numbers of CD8high+(CD57+) cells correlated with previous HCMV infection. By three-colour fluorescence analysis, the CD8high+(CD57 +) population expressed T cell markers such as CD3 and CD5, and most were αβ T cell receptor (αβ TCR)-positive. A significant proportion also expressed CD71 (transferrin receptor) and MHC class II, although little if any CD25 (IL-2R-p55). Some (≥ 40%) co-expressed CD45RA and CD45RO. The CD8low+ (CD57+) population expressed classical natural killer (NK) cell markers—CD2, CD16 and CD56. The two subsets were also functionally distinct; CD8high+ (CD57+) cells suppressed pokeweed mitogen (PWM)-driven, but not phytohaem-agglutinin (PHA)-driven proliferation and immunoglobulin production; CD8low+ (CD57+) cells exhibited NK cytotoxic activity which was not increased by interferon-alpha (IFN-α). Supernatant from cultured CD8high+ (CD57+) cells suppressed PWM-driven immunoglobulin production, but not proliferation, and this effect was abrogated by physical separation with tissue culture inserts. Thus, a T cell subset expressing activation and memory T cell markers with direct non-specific suppressor activity was present in peripheral blood mononuclear cells (PBMC) of healthy subjects with asymptomatic HCMV infection.
IgM and IgG antibodies to hepatitis C virus in patients with mixed cryoglobulinaemia
Authors: L'ABBATE A , CUTRUPI S , ROGNETTA M , FABIANO C , CRAXI A
Published: July 2010

To assess the relationship between hepatitis C virus (HCV) infection and essential mixed cryoglobulinaemia (EMC), sera from 23 patients with EMC were tested for IgG and IgM antibodies to HCV antigens and for HCV RNA. Quantitative HCV antibody studies were performed on scrum and purified cryoglobulin fractions. HCV antibodies of both IgG and IgM class were found in 22 (96%) patients. Ten of these were also HCV-RNA positives. Higher litres of anti-HCV IgM were present in the 11 patients with evidence of liver damage. Anti-HCV IgG antibodies were shown to be concentrated in the IgG fraction of cryoglobulins in all eight patients studied. These results strongly suggest a role for HCV in the pathogenesis of EMC.
DistinctH-2complex control of mortality, and immune responses to tuberculosis infection in virgin and BCG-vaccinated mice
Authors: APT A S , AVDIENKO V G , NIKONENKO B V , KRAMNIK I B , MOROZ A M , SKAMENE E
Published: July 2010

We have studied the impact of distinct haplotypes and of difierent alleles at specific H-2 loci on: (i) the susceptibility to lethal form of experimental tuberculosis; (ii) the level of DTH to mycobacterial antigens: (iii) the efficacy of vaccination with bacille Calmette-Guerin (BCG); and (iv) the IgG production and T cell proliferative response to H37Rv antigens. On the basis of median survival lime (MST) following primary inoculation with lethal dose of Mycobacterium tuberculosis, susceptibility to infection associated with I-Ab and Db alleles. host resistance associated with I-Ak and Dd alleles. Mice bearing a disease-resistant phenotype also developed a vigorous DTH response. Vaccination with BCG before H37Rv infection significantly prolonged the survival time of both resistant and susceptible animals, except in B10.M (H-2f) mice. The latter exhibited intermediate resistance to infection before but slight decrease in the MST following a high-dose BCG vaccination. Distinct H-2 regulation of susceptibility to lethal infection and of BCG vaccination efficacy was confirmed in another relatively resistant H-2f-bearing strain A.CA, in which mortality occurred more rapidly in vaccinated compared with primarily infected animals. The expression of the H-2f haplotype was associated with a low DTH response to tuberculin following vaccination and subsequent lethal infection. The lack of BCG protection against Myco. tuberculosis challenge in B10.M mice associated with the high litre of specific IgG. In addition, these mice exhibited a unique ability to respond to 65-kD antigen by both IgG synthesis and T cell proliferation.