bluuebunny/crossref_metadata_2025 huggingface dataset:
A persistent T cell expansion in the peripheral blood of a normal adult male: a new clinical entity?
Authors: GRUNEWALD J , JEDDI-TEHRANI M , DERSIMONIAN H , ANDERSSON R , WIGZELL H
Published: August 2010

A dramatic and persistent T cell expansion in a healthy adult male was initially identified, using anti-T cell receptor for antigen (TCR)-specific MoAbs. The expanded T cells were found to be expressing TCR containing Vα 12.1 and V β 5.2, and they composed approximately one third of all the CD8+ T cells. The cells were shown to be not only non-activated (HLA-DR−, IL-2R−) but also of ‘virgin’ cell type (CD45RA+/CD45RO−) and they persisted over the observation period of more than one and a half years. Various T and B cell markers, and all other laboratory and physical parameters analysed, were normal. The expanded CD8+ T cells were further characterized by polymerase chain reaction (PCR) amplification, using Vβ- and Cβ-specific primers, followed by hybridization with Jβ-specific probes. Close to 90% of the Vα 12.1+ Vβ 5.2+ T cells were found to utilize the Jβ 2.5 gene segment, thus strongly suggesting the expanded T cells to be monoclonal. The condition may constitute a T cell counterpart to ‘monoclonal gammopathy of undetermined significance’ (MGUS), and by analogy we suggest it should be designated ‘monoclonal T cell expansion of undetermined significance’ (MTUS).
Human MoAbs produced from normal, HIV-1-negative donors and specific for glycoprotein gp 120 of the HIV-1 envelope
Authors: OHLIN M , HINKULA J , BROLIDEN P-A , GRUNOW R , BORREBAECK C A K , WAHREN B
Published: August 2010

Human MoAbs of IgM class were developed against three regions of the HIV-1 envelope. Uninfected donor lymphocytes were immunized in vitro with recombinant protein pBI. Four out of five antibodies were directed to different parts of the V3 region, which contains a major neutralizing site. Two out of these antibodies were directed to more than one amino acid sequence, indicating reactivity to discontinuous sites. Two of the human MoAbs inhibited viral spread between cells in tissue culture, interpreted as reactivities to conserved amino acid sequences exposed during viral maturation. No MoAb neutralized virus, which may be explained by the relatively low avidity of the antibodies. One MoAb was directed to a region containing amino acids participating in CD4 binding. This technique appears to allow formation of antibodies with fine specificities other than those obtained in infected hosts.
Bispecific anti-human red blood Rhesus-D antigen × anti FcγRI targeted antibody-dependent cell-mediated cytotoxicity and phagocytosis by mononuclear leucocytes
Authors: DERAMOUDT F-X , GILARD C , LEPINE N , ALONSO J-M , ROMET-LEMONNE J-L
Published: August 2010

The Fc receptor mediated antibody-dependent cell-mediated cytotoxicity (ADCC) and phagocytosis induced by bispecific antibody (BsAb) to the high-affinity Fc receptor for IgG (FcγRI) and to human red blood group antigen RhD were studied in vitro, using human mononuclear leucocytes as effector cells. The results were compared with those obtained by using a human monoclonal IgGI anti-RhD used alone and a reference human polyclonal anti-RhD antibody. The effect of non-specific human IgG on FcR-mediated functions by mononuclear leucocytes was checked. The results demonstrate that BsAb presents a high resistance of Fc-mediated function to blockade by non-specific human IgG compared with that of both polyclonal and monoclonal anti-RhD antibodies. These results further encourage possible clinical application of bispecfic antibody in passive immunotherapy.
Decreased polymorphonuclear leucocyte chemotactic response to leukotriene B4 in cystic fibrosis
Authors: LAWRENCE R H , SORRELL T C
Published: August 2010

Evidence that leukotriene B4 (LTB4) is a significant inflammatory mediator in chronic pseudomonal respiratory disease was sought in adolescents and young adults with cystic fibrosis. Specific chemotaxis of peripheral blood polymorphonuclear leucocytes (PMN) was used as an indirect measure of remote in vivo exposure to LTB4. PMN from 17 patients showed a significant decrease in chemotaxis to 10−7-10−9m LTB4, but normal responses to 10−8mn-formyl-methionyl-leucyl-phenylalanine and 4 mg/ml casein, when compared with 17 healthy age- and sex-matched controls. This result is consistent with chronic production of LTB4, and specific deactivation of circulating PMN receptors for LTB4 in patients with cystic fibrosis. Pharmacologic inhibition of LTB4 production in vivo may help elucidate its role in the pathogenesis of lung damage in cystic fibrosis.
Characterization of anti-interferon-α antibodies appearing during recombinant interferon-α2a treatment
Authors: RÖNNBLOM L E , TIENSUU JANSONJ E , PERERS A , ÖBERG K E , ALM G V
Published: August 2010

Patients with malignant midgut carcinoid tumours received recombinant interferon-alpha2a (rIFN-α2a) or rIFN-α2a and chemotherapy (streptozocin and doxorubicin) for 6 months, and then rIFN-α2a alone. Antibodies, mainly of IgG type, binding to rIFN-α2a developed in nine of 22 patients (41%), as determined by immunoassay. In seven patients, antibodies also neutralized the biologic (anti-viral) activity of rIFN-α2a. Anti-IFN-α2a antibodies were equally frequent in both sexes and treatment groups, but were not observed in those patients (n=8) that had previously received other types of IFN. Antibodies appeared after a median of 6 months of rIFN-α2a treatment and had a median duration of 6 months. The anti-IFN-α2a antibody titres declined with time with no obvious relation to change of therapy, also during continued IFN-α2a treatment. High titres of neutralizing antibodies appeared to impair anti-tumoural effects in individual potential responders. Anti-IFN-α2a antibodies further examined in six patients bound to native IFN-α subtypes present in both allogeneic and autologous leucocyte IFN-α. Such autoantibodies neutralized the biologic activity of autologous IFN-α in two patients, and in a third were partially neutralizing.
Protective effects of cyclophosphamide, cyclosporin A and FK506 against antigen-induced lung eosinophilia in guinea-pigs
Authors: NORRIS A A , JACKSON D M , EADY R P
Published: August 2010

A close association has been recognized between activated T cells and eosinophils in asthma, albeit circumstantial. The present study attempted to investigate this relationship in an animal model of lung eosinophilia using the new generation of T cell-selective immunosuppressants, cyclosporin A and FK506, compared with the myelotoxic immunosuppressive agent cyclophosphamide. Antigen challenge of ovalbumin-sensitized guinea-pigs resulted in a lung eosinophilia which was assessed by bronchoalveolar lavage. All three agents caused a marked suppression of lung eosinophilia at 24 h post-challenge when the compounds were administered at the time of sensitization but not when administered for 3 days before lavage. However, the lung eosinophilia at 72 h post-challenge was reduced significantly by FK506 and by cyclophosphamide, but not by cyclosporin A, when the drugs were administered for 3 days, before lavage. These results strongly suggest the involvement of T cells in antigen-induced late phase (72 h) eosinophilia in guinea-pigs but not at 24 h. The effects of cyclophosphamide were always associated with a reduction in circulating white cell counts, whereas cyclosporin A and FK506 showed no myelotoxic properties. These results suggest the potential therapeutic use of selective, non-cytotoxic immunosuppressive agents in asthma.
Increased LAK activity against HIV-infected cell lines in HIV-1+ individuals
Authors: GRYLLIS C , WAINBERG M A , BENTWICH Z , GORNITSKY M , BRENNER B G
Published: August 2010

The role of natural killer (NK) cells and their inducible counterparts, lymphokine-activated killer (LAK) cells in AIDS with regard to HIV-1 viral immunosurveillance and the control of secondary opportunistic disease has yet to be established. In this study, we have demonstrated that LAK cells derived from all HIV-1+ groups showed striking increases in their capacity to lyse HIV-1-infected U-937 cells relative to their uninfected U-937 counterparts. Surprisingly, similarly derived LAK cells from healthy seronegative controls showed no differences in their lysis of HIV-1-infected versus uninfected U-937 cells. The differential ability of LAK effectors from seropositive donors to lyse HIV-1-infected targets was demonstrable using a number of U-937 subclones and their HIV-1-infected counterparts. Again, no differences in LAK cell-mediated lysis of HIV-1-infected and uninfected U-937 subclones were observed in seronegative individuals. Our findings that HIV-1+ individuals show selective expansion of non-MHC restricted. HIV-1-directed cytotoxic LAK cells indicate that natural immunity may indeed play a role in HIV-1 viral immunosurveillance.
Regional cellular responses to intraperitoneal infection
Authors: GALANDIUK S , APPEL S , PIETSCH J , OLDFATHER J , POLK H C
Published: August 2010

Leucocytes in different body compartments were examined in a murine intra-abdominal abscess model: (i) peripheral blood: (ii) peritoneal exudate; (iii) abscess wall; and (iv) abscess pus, with respect to their phagocytic ability, CR3 expression and la antigen expression (murine MHC antigen). The ability of serum or supernatant in each compartment to opsonize Staphylococcus aureus was also determined. Mice responded to caecal ligation and puncture (CLP) with systemic monocytosis, an increased amount of opsonins in peritoneal fluid and an increased phagocytosis in pus leucocytes. This increased phagocytic ability and CR3 expression both subsequently decreased. Monocyte/macrophage la expression was reduced in all measured compartments over time. In contrast, sham-operated animals, without an intra-abdominal abscess, responded to operation with an even greater increase in peritoneal exudate phagocytic ability and in monocyte/macrophage Ia expression in all compartments which was sustained beyond 1 week. Decreased peritoneal exudate cell and peripheral blood Ia was still present after 28 days in CLP animals, compared with both sham and normal animals (P < 0·05, p < 0·02, respectively). For all parameters, changes observed in peripheral blood did not reflect those present near the site of infection. Proper understanding of local or regional infection must take into account and ultimately alter these generally unappreciated changes in and about the actual site of infection.
The pattern of mycobacterial antigen recognition in sera from Mantoux-negative individuals is essentially unaffected by bacilli Calmette-Guérin (BCG) vaccination in either South India or London
Authors: DAS S , CHENG S H , LOWRIE D B , WALKER K B , MITCHISON D A , VALLISHAYEE R S , NARAYANAN P R
Published: August 2010

Paired sera were obtained before and 8 weeks after routine BCG vaccination from 20 PPD-S Mantoux-negative individuals who were living adjacent to the Chingleput BCG vaccine trial area in Tamil Nadu and from seven Mantoux-negative school-children in London. UK. Most subjects became Mantoux-positive after vaccination. In ELISA tests against soluble extracts of BCG or Mycobacterium tuberculosis H37Rv or against PPD-S, pre-vaccination antibody titres of South Indian subjects were about twice those of British subjects but there was no increase in titre of antibodies after vaccination of either population. Western blotting showed that even before vaccination, and even in British subjects, antibodies were present that recognized numerous antigenic components in extracts of BCG and M. tuberculosis. There was no consistent difference between band patterns with South Indian and British subjects and any effect of vaccination on the patterns was minimal.
Prevalence of anti-Fab antibodies in patients with autoimmune and infectious diseases
Authors: HASSAN J , FEIGHERY C , BRESNIHAN B , WHELAN A
Published: August 2010

Sensitive ELISA were devised to examine the specificity of circulating IgM and IgA autoantibodies for whole human IgG, Fe and Fab fragments of human IgG. Sera from patients with autoimmune and infectious conditions such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), tuberculosis (TB), infectious mononucleosis (IM) and cystic fibrosis (CF) were studied. Results of the ELISA assays using whole human IgG as antigen revealed that a proportion of patients in each of the groups studied had circulating IgM and IgA rheumatoid factors (RF). Fifteen normal individuals studied were negative. In the latex positive RA group, IgM RF and IgA RF had primarily anti-Fc reactivity (100% and 93% respectively), although 3/15 patients also showed IgM anti-Fab reactivity and one patient had high IgA anti-Fab activity. Patients with SLE and TB who had detectable RF levels also revealed predominantly anti-Fc specificity. In contrast, examination of 25 patients with IM showed positivity for IgM RF activity in 8% of patients using whole IgG as antigen, 24% positivity using purified Fc fragments as antigen and 45% positivity when plates were coated with Fab fragments. Similarly, a large number of CF patients (54%) also showed predominantly IgM anti-Fab activity. Of interest, 69% of the CF patients who were all studied at the time of bacterial infection had detectable IgA RF levels, with 46% of these patients showing both IgA anti-Fc and anti-Fab activity. These findings suggest that autoantibody specificities in autoimmune and infectious diseases are different.
A recombinant topoisomerase I ELISA: screening for IgG, IgM and IgA anti-topo I autoantibodies in human sera
Authors: VERHEIJEN R , de JONG B A W , van VENROOIJ W J
Published: August 2010

An ELISA for the detection of anti-topoisomerase I autoantibodies in sera from patients with suspected or manifest rheumatic diseases is described. The antigen source used in this assay consists of a recombinant protein containing the last 695 C-terminal amino acid residues of human topoisomerase I (topo I), The sensitivity of the assay was 61%, while the specificity was more than 98%. Using this ELISA, 47 sera from scleroderma patients and immunopositive for anti-topo I antibodies, were screened for the presence of the isotypes IgG, IgA and IgM to topo I. Our finding that relatively high levels of IgA antibodies to topo I are present in most of the sera tested is consistent with the results of Hildebrandt et al. [1]. In addition, it is demonstrated that the IgG and IgA antibodies in a serum may recognize different epitope regions on the topo I polypeptide.
Expression of CD56 (NKH-1) differentiation antigen in human thyroid epithelium
Authors: ZEROMSKI J , BAGNASCO M , PAOLIERI F , DWORACKI G
Published: August 2010

Leu-19 (CD56) MoAb is well known to recognize gp220 expressed on natural killer (NK) cells and is widely used as a NK cell marker. The expression of CD56 antigen was tested by means of sensitive alkaline phosphatase-anti-alkaline phosphatase (APAAP) immunohistochemical technique and the above mentioned MoAb as a primary antibody, on frozen sections of various fresh human tissues. Out of 11 organs examined only thyroid gland provided a distinct reaction confined to cell membranes of epithelial follicular cells. The reaction had a diffuse pattern in cases of Graves' disease and colloidal goitre while in Hashimoto's thyroiditis presented as a focal pattern. Other anti-NK cell MoAbs such as VD4 (CD16) and Leu-7 (CD57) reacted only with single cells of thyroid stroma. The results of APAAP staining were confirmed by the cytofluorimetric assessment of isolated thyroid cells. It is speculated that CD56 expression on thyroid cells may have a functional significance, perhaps related to neural–endocrine interactions.
Significance of C3 nephritic factor (C3NeF) in non-hypocomplementaemic serum with membranoproliferative glomerulonephritis (MPGN)
Authors: OHI H , WATANABE S , FUJITA T , YASUGI T
Published: August 2010

C3NeF is an autoantibody of C3 convertase (C3bBb) and is often detected in the serum of hypocomplementaemic MPGN patients. Serum samples from 104 non-hypocomplementaemic MPGN patients (C3NeF) were studied. C3NeF, which cannot activate the alternative pathway, was found in the sera of 6 patients. We examined the C3NeF in purified IgG from five of the non-hypocomplementaemic serum samples (non-hypo C3NeF) and four hypocomplementaemic serum samples (hypocomplementaemic C3NeF) to determine why C3NeF does not induce C3 splitting and hypocomplementaemia. Purified IgG from non-hypo C3NeF stabilized EAC4b3bBb cells in a manner similar to IgG from hypocomplementaemic C3NeF in EDTA gelatin veronal buffer. However, the non-hypo C3NeF IgG did not stabilize C3 convertase (EAC4b3bBb cells) in the presence of control proteins (factors H and I), whereas the hypocomplementaemic C3NeF IgG did. The C3NeF in the hypocomplementaemic serum displayed two characteristics: (i) inhibition of intrinsic decay of Ce convertase (C3bBb); and (ii) inhibition of extrinsic decay by factors H and I. Although the C3NeF in the non-hypocomplementaemic sera did inhibit the intrinsic decay in a manner similar to the hypocomplementaemic C3NeF IgG, it did not inhibit the extrinsic decay. Due to the different characteristics of hypocomplementaemic C3NeF and non-hypo C3NeF in the serum samples, the non-hypo C3NeF did not activate C3. Therefore, we conclude that C3NeF exhibits a heterogeneity which is very important in relation to the pathogenesis of MPGN.
C7 M/N protein polymorphism typing applied to inherited deficiencies of human complement proteins C6 and C7
Authors: WÜRZNER R , RANCE N , POTTER P C , HENDRICKS M L , LACHMANN P J , ORREN A
Published: August 2010

C7 M/N typing, the determination of the complement component C7 M/N phenotypes, was successfully used in family studies to trace haplotypes bearing C7 deficiency genes. Furthermore, it was shown to be preferable to CT allotyping based on isoelectric focusing (IEF) since it distinguishes two common alleles (C7*M and C7*N), whereas one common C7IEF allele (C7*I) predominates in most populations. It is also the more sensitive method, as it enabled detection of very low amounts of abnormal C7 molecules in the third generation of a combined subtotal C6/C7-deficient subject and thus confirmed that this partial deficiency gene is not silent in heterozygotes. In this respect C7 M/N typing is even more informative than DNA restriction fragment length polymorphism typing which will assess the presence but not necessarily the functional status of a gene. C6 and C7 genes are tightly linked and therefore C7 M/N typing was also applied to tracing C6 deficiency genes in families.C6/C7 haplotype analysis of South African C6-deficient tC6Q0) subjects revealed a strong allelic association of C6*Q0 and C7*M.
Soluble forms of membrane cofactor protein (CD46, MCP) are present in plasma, tears, and seminal fluid in normal subjects
Authors: HARA T , KURIYAMA S , KIYOHARA H , NAGASE Y , MATSUMOTO M , SEYA T
Published: August 2010

We have established an ELISA for determination of membrane cofactor protein (MCP, CD46) both solubilized from cell membranes and released in body fluids. In this assay, mouse MoAbs against MCP, M177 and M160 whose epitopes were different, were used as capture and detection antibodies, respectively. The NP-40 concentration in samples for MCP to be measured must be less than 0·05%. The detection limit of this MCP assay was 0·5 ng. The assay was used to quantify solubilized membrane MCP, and soluble MCP in normal human plasma, serum, urine, saliva, tears, and seminal fluid, and culture media of tumour cell lines. Soluble MCP was barely detected in the conditioned media of the cell lines. The levels of sMCP in plasma and serum were 10–60 ng/ml and that in tears, 0–50 ng/ml. Seminal fluid contained about 10-fold more soluble MCP than serum. Soluble MCP was not detectable by this assay in the other body fluids, suggesting that their MCP levels were less than the detection limit, if any.
Abnormal immunoglobulin G subclass production in response to keyhole limpet haemocyanin in atopic patients
Authors: FALCONER A E , FRIEDMANN P S , BIRD P , CALVERT J E
Published: August 2010

A proportion of patients with atopic dermatitis have elevated serum levels of IgG4. In order to investigate further this abnormality of IgG subclass production, atopic patients were immunized with the protein antigen keyhole limpet haemocyanin (KLH), and IgG subclass responses following primary and secondary immunization were analysed. In the primary response, titres of IgG1, 2 and 3 antibodies were lower in the atopic patients than in the controls. In contrast, titres of IgG4 were much higher for the patient group. In both patients and controls, the kinetics of IgG4 antibody production following the initial immunization with KLH showed a slow rise reaching a peak at 30 weeks. This time course indicated that the high IgG4 response was unlikely to be due to previous exposure of the patients to a cross-reacting antigen. A higher proportion of IgG4 was also seen in the atopic patients following secondary immunization; indeed, IgG4 was the major subclass in the secondary response in the patient group. In the controls, but not in the patients, titres of IgG4 anti-KLH correlated with total serum levels of IgG4, and some of the highest IgG4 antibody responses were detected in atopic patients whose serum IgG4 concentration was in the normal range. The results suggest that raised serum levels of IgG4 in atopy may reflect abnormal isotype regulation in response to protein antigens.
Production of reactive nitrogen intermediates by bone marrow-derived macrophages on treatment with cisplatinin vitro
Authors: SODHI A , SURESH A
Published: August 2010

L929 culture medium (a source of macrophage colony stimulating factor (M-CSF) or recombinant granulocyte-macrophage colony stimulating factor (rGM-CSF)-derived bone marrow macrophages treated with cisplatin or lipopolysaccharide (LPS) (10 μ/ml) were effective in the production of L-arginine-dependent reactive nitrogen intermediates (RNI) and generation of tumouricidal activity. The abilities of RNI secretion and related tumouricidal activity against P 815 mastocytoma cells were compared. These parameters were found to be closely correlated in various experiments, RNI secretion and generation of bone marrow macrophage-mediated tumouricidal activity were significantly inhibited by l-n-monomethyl arginine (l-NMMA), a specific inhibitor of the L-arginine pathway, but l-NMMA did not inhibit macrophage-mediated killing of tumour necrosis factor (TNF)-sensitive Wehi cells, suggesting that activated macrophages exhibit at least two cytolytic mechanisms, one by L-arginine-dependent nitric oxide pathway and another by TNF-mediated killing. The present findings suggest that the mechanism of tumour cell killing by activated macrophages may differ, depending on the tumour cell type, and reactive nitrogen intermediates play a major role in cisplatin-mediated activation of bone marrow-derived macrophages.
A gp120 HIV peptide with high similarity to HLA class II β chains enhances PPD-specific and autoreactive T cell activation
Authors: PUGLIESE O , VIORA M , CAMPONESCHI B , CORDIALI FEI P , CAPRILLI F , CHERSI A , EVANGELISTA M , DI MASSIMO A M , COLIZZI V
Published: August 2010

The recent report that anli-gp120 antibodies can be induced by allogeneic stimuli in experimental animals in the absence of HIV, has focused attention on the structural similarities between gp120 and MHC. Here we report that some HIV+ individuals develop antibodies which similarly react with the gp120 HIV sequence (aa 254–263) and with the HLA-DR β chains (aa 142–151). As these two peptides share a high level of similarity, we have investigated the role of this gp120 region on HLA class II mediated T cell recognition. The synthetic peptide corresponding to the gp120 HIV sequence aa 254–263 has been tested on T cell line (TCL) activation. Both the PPD-specific and the self-HLA reactive TCL proliferation increased in the presence of this peptide. Prepulsing experiments indicate that this enhancing effect carried out by HIV peptide is exerted at the level of antigen presentation. Moreover, the specificity of this interaction is supported by the fact that a MoAb specific for this HIV peptide blocked the autoreactive TCL proliferation, similarly to the inhibition carried out by anti-class II antibody. These data support the hypothesis that the functional homology between the HIV peptide and the HLA β chain described may be involved in the pathogenesis of AIDS.
Immune response to hepatitis B virus surface antigen peptides during HBV infection
Authors: MISHRA A , DURGAPAL H , MANIVEL V , ACHARYA S K , Rao K V S , Panda S K
Published: August 2010

Antibody responses of patients with acute (n = 73), fulminant (n = 30) and chronic (n = 51) hepatitis B virus (HBV) infection as well as recovered individuals (n = 7) were studied against three synthetic peptides, Pre-S1 amino acids (aa. 12–32), Pre-S2 amino acids (aa. 120–145), and S amino acids (aa. 124–147) of the envelope region (HBsAg). T cell blastogenic response was investigated in a proportion of the patients (27 acute, nine fulminant, 13 chronic hepatitis and seven recovered individuals) along with seven HBV vaccinated and three normal individuals. The presence of T cell response against S peptide was observed in all the cases (9/9, 100%) during early acute hepatitis. This was suppressed during late stages (8/18, 44%) followed by partial reversal during recovery (5/7, 71 %). T cell response and antibodies to Pre-S1 and Pre-S2 peptides were present only in one-third of the patients throughout these periods. The T cell blastogenic response as well as antibody reactivity against these peptides were absent and minimal in chronic hepatitis. Immune response against envelope protein appears to play a major role in acute hepatic injury due to HBV infection and help in virus clearance.
Phenotypic analysis of lymphocyte populations in the lungs and regional lymphoid tissue of sheep naturally infected with maedi visna virus
Authors: WATT N J , MacINTYRE N , COLLIE D , SARGAN D , McCONNELL I
Published: August 2010

We have analysed the phenotype of lymphocytes in lung and regional lymph node of symptomatic and asymptomatic sheep infected with the ovine lentivirus, maedi visna virus (MVV). Compared to equilavent tissues from age-matched, non-infected controls, MVV-infected sheep show increased numbers of lymphocytes in the lung, both in the bronchus-associated lymphoid tissue (BALT) and in the alveolar septae. Both CD8+ and CD4+ T lymphocyte numbers in alveolar septae were increased, particularly in animals with clinical respiratory disease. The ratio of CD8+ to CD4+ lymphocytes was similar to that in normal lung. In both MVV-infected and uninfected animals a high proportion of pulmonary lymphocytes, particularly in the alveolar septae, did not express the CD5 antigen, suggesting that they were activated. The number of activated cells was higher in infected sheep. Variable numbers of alveolar macrophages containing MVV-core protein were present in alveolar lumina, the majority of positive cells showing morphological evidence of activation. In regional lymphoid tissue there were increased numbers of CD8 + and γδ expressing T cells in lymphoid follicles and germinal centres of infected animals. The specificity of these cells is unknown and we could find no evidence for the presence of cells productively infected with the virus in these structures. This study shows that activated T lymphocytes, particularly of the CD8 subset, play a major part in the pathogenesis of MVV-induced pulmonary and regional lymph node lesions.
Cellular changes in the bronchoalveolar lavage (BAL) of pigs, following immunization by the enteral or respiratory route
Authors: DELVENTHAL S , HENSEL A , PETZOLDT K , PABST R
Published: August 2010

Normal young pigs were immunized by the oral or aerogenic route with the viable or inactivated lung-pathogenic bacterium Actinobacillus (Haemophilus) pleuropneumoniae. Three weeks later the cellular composition as well as the lymphocyte subset composition of the bronchoalveolar space were examined by BAL. Lymphocytes in the lavage increased significantly, including CD4+ and CD8+ T cells. After oral immunization a dramatic increase of plasma cells and lymphoid blasts was found. Among immunoglobulin-positive lymphocytes IgG+ cells showed the most pronounced increase. For most lymphocyte subsets there was no difference between viable and inactivated bacteria. Oral immunization with a lung-pathogenic bacterium results in increased numbers of lymphocytes in the bronchoalveolar space and might play a critical role in protection against lower respiratory tract infections.
Suppression of spontaneous murine lupus by inducing graft-versus-host reaction with CD8+ cells
Authors: ITO S , UENO M , NISHI S , ARAKAWA M , FUJIWARA M
Published: August 2010

Splenic CD8+ (Lyt-2+) cells of C57BL/6 mice were injected into semiallogeneic (NZB × BXSB)F1 mice, which spontaneously develop lupus nephritis, in order to examine whether the disease was somehow modified by the occurrence of graft-versus-host reaction. The development of lupus nephritis in the F1 recipients was strongly inhibited and immunopathological parameters such as anti-DNA antibodies, circulating immune complexes (CIC) and splenic immunoglobulin-producing cells (IgPC) were markedly reduced. The injection of CD4+ (L3T4+)Tcells into F1 recipients did not result in similar effects. These findings suggest that the development of autoimmune disease could be ameliorated by CD8+ cells responding to MHC class I antigens. The significance of the data is discussed in terms of the treatment of autoimmune diseases.
Circulating antibodies to heat-shock protein 60 in Crohn's disease and ulcerative colitis
Authors: STEVENS T R J , WINROW V R , BLAKE D R , RAMPTON D S
Published: August 2010

Heat-shock proteins (HSPs) are highly conserved immunogenic intracellular molecules that are induced by inflammatory mediators and may induce autoimmune phenomena in vivo. We have recently demonstrated the increased expression of HSP-60 in the colonocytes of patients with ulcerative colitis. To study further the role of HSP-60 in inflammatory bowel disease, we have now measured antibodies to recombinant mycobacterial HSP-65 (a member of the HSP-60 family) in patients with Crohn's disease, ulcerative colitis, healthy volunteers and, as disease controls, patients with confirmed bacterial diarrhoea. In comparison with healthy controls (n = 20; median level of 89 ELISA units; range 24–292), serum IgA HSP-60 antibodies were elevated in Crohn's disease (n = 21; 157; 57–364; P<0·05) and active ulcerative colitis (n= 16; 188; 58–373; P<0·01) but not bacterial diarrhoea (n= 10; 106; 51–285). Increased IgA HSP-60 antibody levels in patients with inflammatory bowel disease may occur as the result of HSP release from injured gut epithelium; alternatively, increased intestinal permeability could facilitate mucosal access of luminal antigens and the generation of cross-reactive anti-bacterial HSP antibodies.
Leukotriene B4-induced hyperadhesiveness of endothelial cells for neutrophils: relation to CD54
Authors: PALMBLAD J E W , LERNER R
Published: August 2010

Leukotriene B4 (LTB4) induced an in vitro transient state of hyperadhesiveness in cultured human umbilical vein endothelial cells (HUVEC), leading to a 2·2-fold increase in the binding of neutrophil granulocytes (PMN), which was less than that conferred by platelet activating factor (PAF) though more than thrombin did (3·4- or 2·0-fold increases, respectively). This study concerns the role of the adhesive molecules CD 18 and CD54 for the LTB4- (as well as thrombin- and PAF-) induced endothelial hyperadhesiveness. The MoAbs 60·3 (to the CD18 molecule on PMN) and 84 H 10 (to one epitope of CD54 on the HUVEC) blocked the adherence of PMN to LTB4-treated HUVEC, whereas MoAb LB-2 (directed at another CD54 epitope) failed to do so. MoAb 84 H 10 blocked 43% of the thrombin-induccd hyperadhesiveness, whereas the PAF response was unaffected. Thus, LTB4-induced HUVEC hyperadhesiveness may therefore be related to a specific domain on the CD54 (or on an antigenically related molecule) as well as being dependent on CD 18, whereas the involvement of CD54 was much less or non-existent for the thrombin and PAF responses, respectively.
Passive immunization against tumour necrosis factor-alpha (TNF-α) and IL-lβ protects from LPS enhancing glomerular injury in nephrotoxic nephritis in rats
Authors: KARKAR A M , KOSHINO Y , CASHMAN S J , DASH A C , BONNEFOY J , MEAGER A , REES A J
Published: August 2010

Glomerular injury caused by injection of heterologous anti-glomerular basement membrane antibodies (anti-GBM Ab) is increased in rats pretreated with small doses of bacterial lipopolysaccharide (LPS). We have investigated the involvement of tumour necrosis factor-alpha (TNF-α), IL-lα and IL-1 β in this phenomenon by passive immunization against these cytokines. Anti-TNF-α or anti-IL-1β antibodies given 1·5 h before the induction of nephritis significantly decreased injury in this model, whether assessed by the magnitude of albuminuria, the prevalence of glomerular capillary thrombi or the intensity of glomerular neutrophil infiltrate. Albuminuria in anti-GBM Ab alone was 11 ± 3, LPS/anti-GBM Ab 87±22, and anti-TNF-α antibodies/LPS/anti-GBM Ab 21 ±6 mg/24 h (mean ± s.e.) P< 0·05. Passive immunization with antibodies to IL-1 β had a similar effect (anti-GBM Ab, 0·6±01, LPS/anti-GBM Ab, 92±19, anti-IL-1β antibodies/LPS/anti-GBM Ab 39 ± 8 mg/24 h, P<0·05). The prevalence of glomerular capillary thrombi was also reduced significantly by these treatments; from 22 ± 5% to 4± 1% in the case of anti-TNF-α antibodies and 28±5% to 13±4% with anti-IL-1β antibodies. Similarly, the glomerular neutrophil infiltrate was also reduced by these treatments; from 42±3 to 25±1 in the case of anti-TNF-α and 47± 2 to 30± 1 with anti-IL-1α antibodies. In contrast, passive immunization against IL-1α had no effect on either albumin excretion (4±3, 83±22 and 77±24 mg/24 h), glomerular capillary thrombi (2±1; 19±5 and 16±3) or glomerular neutrophil infiltrate (22± 3; 47± 5 and 48± 5 from the three groups respectively). These results demonstrate that enhanced antibody mediated injury in the kidney is modulated by TNF-α and IL-14bT but not by IL-lα.
Analysis of lymphocyte phenotypes in cord blood from early gestation fetuses
Authors: PEAKMAN M , BUGGINS A G S , NICOLAIDES K H , LAYTON D M , VERGANI D
Published: August 2010

Using cord blood samples obtained from Fetuses between 16 and 40 weeks gestation, we have used a lysed whole blood flow cytometric technique to study the natural history of lymphocyte phenotypes known to be highly represented in cord blood at birth. The majority (51·0 ± 14·7%) of lymphocytes expressed CD45RA, a marker of ‘virgin’ cells and there was a correIation between increasing percentages of CD45R A+ lymphocytes and gestational age (r = 0·44, P<0·01). Few cells (8·5 ± 4·2%) expressed the CD45RO marker of primed lymphocytes and very few (1·0 ± 0·7%) co-expressed CD45RA and RO, indicating little traffic between the two maturation markers. The percentage of B lymphocytes co-expressing CDS was high in the fetal circuIation (55·5±10·5%) compared with healthy adults (23·2 ± 14·3%; P<0·00001) and the level of CDS+ B cells declined with gestational age in an exponential manner (r=−0·45, P<0·05). Similarly, levels of T lymphocytes expressing the γδ T cell receptor (TCR) declined exponentially (r=−0·59, P<0·005). These results demonstrate that lymphocytes remain almost entirely unprimed before birth. In addition, CD5+ B lymphocytes and TCR-γδ+ T lymphocytes decline exponentially towards birth, in a manner suggesting that they may be seeding peripheral sites such as the spleen, skin and mucosae.
Changes in natural immunity during the course of HIV-1 infection
Authors: BRENNER B G , GRYLLIS C , GORNITSKY M , WAINBERG M A
Published: August 2010

The role of natural killer (NK) and lymphokine-activated killer (LAK) cell-mediated cytotoxiciry in AIDS has yet to be established. The objective of this study was to determine inducible LAK cell responses at different stages of HIV-1 infection, and specifically to establish the participation of CD8 lymphocytes in these responses. Peripheral blood lymphocytes (PBL) were isolated from healthy seronegative (CDC-0) subjects and HIV-1 individuals who were clinically asymptomatic (Centre for Disease Control group 2, CDC-2) or symptomatic (CDC-4) with regard to secondary opportunistic infection (OI). LAK cells were generated upon incubation of PBL with IL-2 and their cytolysis of K562 and U-937 targets was determined using chromium release assays. The role of CD8+ lymphocytes as progenitors and effectors of these LAK cell responses was determined by immunomagnetic depletion of CD8+ cells from precursor PBL and LAK cells, respectively. LAK cell-mediated cytotoxicities in HIV-1-infected individuals were reduced compared with scronegativc controls without any corresponding changes in the relative proportions of CD56+ (NK) cells among groups. Depletions of CD8+ subsets from either PBL or LAK cells dramatically reduced total LAK cytotoxic responses and LAK activities per unit CD56+ cell in the OI-CDC-2 scropositive population. No corresponding changes in LAK activities in seronegative control or HIV+/OI+ CDC-4 groups were observed. Levels of LAK activity against K562 targets in CDC-0/HIV and CDC-4/HIV+ groups correlated with the percentage of CD56+ LAK cells; corresponding LAK activity in the CDC-2/HIV+ group correlated with the percentage of both CD56+ and CD8+ subsets. These findings suggest that adaptive changes in non-MHC restricted cytotoxic responses occur in HIV-1 individuals at early stages post-HIV infection, before the onset of opportunistic infection.
Elevated serum levels of IL-8 in patients with HIV infection
Authors: MATSUMOTO T , MIIKE T , NELSON R P , TRUDEAU W L , LOCKEY R F , YODOI J
Published: August 2010

Scrum levels of IL-8 were determined in HIV-infected individuals and the results were compared with those for HIV controls. The IL-8 levels were measured by an ELISA with a MoAb and a polyclonal antibody to recombinant IL-8. The means and 95% confidence intervals of IL-8 in sera of 36 HIV-infected individuals and 32 matched controls were 275 and 216 349 pg/ml, and 8 and 4 14 pg/ml, respectively, showing a 34-fold increase in IL-8 in the circulation of HIV-infected individuals. This increase does not appear to be related to the disease state, infection or systemic medical agents. This finding suggests the possible involvement of IL-8 in the pathogenesis of HIV-induced disease.
Ring-infected erythrocyte surface antigen (Pf/155RESA) induces tumour necrosis factor-alpha production
Authors: PICOT S , PEYRON F , DELORON P , BOUDIN C , CHUMPITAZI B , BARBE G , VUILLEZ J P , DONADILLE A , AMBROISE-THOMAS P
Published: August 2010

Cerebral malaria is probably related to an overstimulation of the immune system and the cytokinc network. We have previously demonstrated that tumour necrosis factor (TNF) secretion by human macrophages can be induced by soluble and heat-stable malarial antigens. Indirect evidence from epidemiological and in vitro studies suggests that Pf155/RESA can be considered as a candidate for triggering TNF secretion. Thus we conducted experiments to investigate the relationship between Pf155/RESA and TNF production. The SGE1 strain of Plasmodium falciparum was compared with the P. falciparum FCR3 strain, which does not express Pf155/RESA protein, for ability to induce TNF secretion by normal human macrophages in vitro. Synthetic peptides from the Pf155/RESA antigen ((EENV)4, (EENVEHDA)4, (DDEHVEEPTVA)3), were used in some experiments. TNF levels were measured by an immunoradiometric assay. We observed that the RESA-defective strain induces lower levels of TNF after schizont rupture than the SGE1 strain. Moreover, substantial TNF secretion was detected when macrophages were incubated with all three peptides, maximum levels being obtained with the(EENV)4 peptide. Although previous reports have described TNF-inducing activity of phospholipid from P. falciparum, these findings strengthen the evidence for Pf155/RESA antigens also being involved in TNF production during malaria.
Determination of soluble CD21 as a parameter of B cell activation
Authors: HUEMER H P , LARCHER C , PRODINGER W M , PETZER A L , MITTERER M , FALSER N
Published: August 2010

In this study we established a novel solid-phase immunoassay for CD21 using the time-resolved fluorescence of lanthanide chelates. The capture assay was able to detect concentrations of as low as 100 pg of CD21 antigen per millilitre of sample and was used for quantitative determination of CD21 in lysates of different cell lines as well as in patient serum specimens. CD21 was measured in lysates of tonsils and cell lines of B. T cell and myelomonocyte lineage, and appeared to consist of monomeric antigen under the detergent conditions used. Elevated levels of soluble CD21 were observed in serum of patients with Epstein Barr virus (EBV) infection, a disease known to be associated with polyclonal B cell activation, and in infection with the lymphotropic rubella virus. Significantly increased levels were also found in malignancies which are associated with EBV. In patients with nasopharyngeal carcinoma (NPC), a correlation with the titre of EBV-specific IgA was observed, thus supporting a possible role of soluble CD21 as a marker for disease activity in certain malignancies. Our data suggest that measurement of soluble CD21 could serve as a marker for activation of the immune system and diseases involving the B cell lymphoid system. Possible mechanisms and functions of soluble CD21 are discussed.
Early induction of IL-1 receptor antagonist (IL-1Ra) in infants and children undergoing surgery
Authors: NUALLÁIN E M Ó , PURI P , REEN D J
Published: August 2010

The cytokine response to injury or trauma is of interest in terms of both its mediation of the acute phase response and its possible relation to the immunological depression observed after major surgery. In this study, the production of cytokines IL-1 β, tumour necrosis factor-alpha (TNF-α), IL-6 and the naturally occurring inhibitor of IL-1, IL-1Ra, have been investigated in infants and children undergoing Swenson's pull-through operation for Hirschsprung's disease. Samples of peripheral blood were taken before, during and after surgery for the measurement of cytokines. IL-IRa levels increased significantly (P <0.01) at 2 h after commencement of surgery, with maximal levels for individual patients being attained between 3 h and 5 h (range 7.6–67.9 ng/ml). The mean level of IL-1 Ra was maximal (26.2 ng/ml) at 5 h and returned to baseline levels between 24 h and 72 h. There were no changes observed in the circulating levels of IL-1β in nine out of 11 patients following commencement of surgery. TNF-α levels did not increase in any of the patients studied. IL-6 levels increased significantly (P<0.02) 3 h after commencement of surgery, reaching maximum concentrations at 24 h (range 20–670 pg/ml), with levels falling between 48 h and 72 h. This study demonstrates, in vivo, the independent induction of IL-IRa without a concomitant increase of IL-lβ levels after major surgery. It also shows that IL-1Ra is the earliest cytokine produced in response to surgical stress.
Suppression of anti-erythrocyte autoantibody-producing B cells by a physiological IgG-anti-F(ab')2 antibody and escape from suppression by tumour transformation; a model relevant for the pathogenesis of autoimmune haemolytic anaemia
Authors: TERNESS P , MARX U , SANDILANDS G , ROELCKE D , WELSCHOF M , OPELZ G
Published: August 2010

We showed previously that broadly reactive IgG anti-immunoglobulin autoantibodies produced by rats during the immune response suppress the B cell response. We report here on the effect of a similar human antibody on self-reactive human B cells. IgG anti-F(ab')2 was added to cultures of anti-erythrocyte autoantibody-producing B cells derived from healthy donors. A dose-dependent suppression of the antibody response was obtained (maximum at 1.3 ng IgG/106 cells). This effect was competitively inhibited by F(ab')2γ. Autoimmune haemolytic anaemia can be caused by chronic monoclonal B cell proliferation. To reproduce this condition in vitro we immortalized B cells with Epstein-Barr virus (EBV) and raised a B cell population with anti-erythrocyte autoantibody activity. These cells were electrically fused with CB-F7 tumour cells and an IgGl cold-reactive anti-erythrocyte autoantibody-producing B cell line was established. Surprisingly, the tumour cells were not suppressed by IgG anti-F(ab')2. It is known that anti-immunoglobulins selectively suppress antigen-receptor (AgR)-occupied B cells by a Fcγ-receptor (FeyRemediated mechanism. To occupy their AgR, we preincubated the tumour cells with anti-AgR antibody. In spite of this, their susceptibility to suppression was not restored. As shown by rabbit IgG-sensitized ox erythrocyte (EA)-rosetting, this refractoriness was not due to a loss of FcγR. Our experiments delineate a mechanism of peripheral B cell suppression to autoantigens, and show a way of escape from control relevant for the pathogenesis of autoimmune haemolytic anaemia.
Induction of phospholipid-binding antibodies in mice and rabbits by immunization with human β2 glycoprotein 1 or anticardiolipin antibodies alone
Authors: PIERANGELI S S , HARRIS E N
Published: August 2010

Anticardiolipin (aCL) antibodies are autoantibodies present in high concentrations in patients with the antiphospholipid syndrome (APS), a disorder of recurrent thrombosis and pregnancy loss. What induces aCL antibodies is uncertain, but a recent report suggested that immunization of mice with β2glycoprotein 1 (β2GPI)in Freund's complete adjuvant (FCA) resulted in aCL antibody production in the recipient mice. Since this observation might explain how autoantibodics might be induced by poor immunogens, such as phospholipids, we decided to explore the question further. In our first series of experiments, we found that aCL antibodies were induced in mice by β2GP1 mixed with adjuvants that did not contain lipids (Adju-Prime or aluminium hydroxide). This excluded the possibility that antibody induction occurred because β2GPl formed complexes with lipids in FCA. We also found that aCL antibodies always appeared before anti-β2GPl antibodies, excluding the possibility that aCL antibodies were directed to β2GP1 or were induced by formation of anti-idiotypic antibodies (to anti-β2GPl). In experiments, we found that immunization of mice with human IgG antibodies from patients with the APS (IgG-APS), also induced aCL antibodies. Immunization with pure bovine serum albumin (BSA) did not induce aCL antibodies. We propose that aCL antibodies arc induced by proteins with high avidity for phospholipids. These proteins may be bound to phospholipids when introduced, or may bind circulating phospholipids. so transforming phospholipid molecules into immunogens. Similar mechanisms might explain autoantibody induction to other poor immunogens.