bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunoglobulin and antibody levels in bronchoalveolar lavage fluid from symptomatic and asymptomatic pigeon breeders
Authors: REYNOLDS S P , EDWARDS J H , JONES K P , DAVIES B H
Published: July 2010

Twenty-one symptomatic subjects with pigeon breeders’ lung (PBL) and 10 asymptomatic pigeon breeders, with a similar exposure to pigeon antigens, underwent bronchoalveolar lavage. Total IgG, IgM and IgA in lavage fluid were determined as were specific antibody levels against antigens in pigeon serum and droppings. Results were converted to levels in epithelial lining fluid (ELF) using lavage and serum urea ratios. It was found that symptomatics represent a group that is hyperreactive to pigeon antigens compared with the asymptomatic group with significantly higher IgG, IgM, IgA levels as well as specific antibody levels against pigeon serum and droppings. Paired serum and ELF samples from 12 symptomatic subjects showed significantly elevated IgG, IgM and IgA levels in ELF compared with serum when values were expressed in terms of albumin. This strongly supports the concept of local production of immunoglobulins within the lung after inhaling immunogens as opposed to their diffusion from the vasculature. Results for IgA indicate that any putative protective role for this immunoglobulin is not valid in relation to the prevention of extrinsic allergic alveolitis. Analysis of smoking habits, lung immunoglobulins and response to inhalation challenge confirm the negative influence of smoking on total and functional lung immunoglobulins; however, levels in the ELF of ex-smokers suggest that the effect of smoking is not permanent. Smoking did not prevent responses to inhalation challenge.
Changes in serum immunoglobulin patterns in adults with common variable immunodeficiency
Authors: SELIGMANN M , AUCOUTURIER P , DANON F , PREUD'HOMME J L
Published: July 2010

Striking variations of serum immunoglobulin class and IgG subclass levels were observed in five patients with common variable immunodeficiency. They occurred mainly in untreated patients or, in those patients who received substitutive therapy, could not be merely due to replacement. They result in major changes in the immunoglobulin deficiency patterns, such as a shift from profound hypoimmunoglobulinaemia to IgA/IgG2/lgG4 deficiency or to isolated IgG2 deficiency. These findings have practical implications for the diagnosis and management of patients with primary humoral immunodeficiency.
Altered production of tumour necrosis factors alpha and beta and interferon gamma by HIV-infected individuals
Authors: VYAKARNAM A , MATEAR P , MEAGER A , KELLY G , STANLEY B , WELLER I , BEVERLEY P
Published: July 2010

In vitro studies shows that recombinant tumour necrosis factor (TNF) α and β, and interferon-gamma (IFN-γ) can enhance HIV replication, and peripheral blood mononuclear cells (PBMC) infected with HIV in vitro secrete high levels of the same cytokines. As T cells secrete all three mediators, the capacity of T cell activation signals to trigger cytokine production in PBMC from HIV-infected individuals was investigated as such patients may be immunocompromised. We demonstrate that asymptomatic seropositives in CDC group II/III as well as patients who have progressed to CDC group IV of the disease proliferate efficiently to anti-CD3 antibody, recombinant interleukin-2 (rIL-2), phytohaemagglutinin (PHA), PHA plus phorbol 12, 13 dibutyrate (PMA) but secrete significantly (P>0.05) higher amounts of TNF-α, TNF-β and IFN-γ compared with controls in response to the same stimulants. We also show a difference between group II/III and group IV patients with the latter secreting more TNF-α and IFN-γ. The kinetics of TNF-α and -β and IFN-γ production was stimulus dependent with overall levels varying in time for each stimulus. Furthermore, the kinetics of the response to all three stimulants were altered in seropositives; CDC group II/III and group IV patients secreted higher levels of cytokines over several time points compared to controls. The altered production of these mediators by HIV-infected patients may contribute to disease progression and to the pathogenesis of AIDS.
Characterization of anti-endothelial cell antibodies in systemic lupus erythematosus (SLE)
Authors: VAN DER ZEE J M , SIEGERT C E H , DE VREEDE T A , DAHA M R , BREEDVELD F C
Published: July 2010

IgG anti-endothelial antibodies (AEA), as measured by EL1SA or immunoblotting technique could be delected in serum samples or 56 out of 64 patients with SLE (88%) and mainly occurred in monomeric form. AEA were not cell specific, because the binding reactivity was absorbed partially by both fibroblasts and peripheral blood mononuclear cells. No correlation was found between the presence of AEA and anti-nuclear antibodies. Immunoblotting revealed reactivity of AEA against endothelial antigens ranging in size from 15 to 200 kD. AEA litres were significantly higher in patients with joint or skin abnormalities, compared with patients without these abnormalities. A significant correlation was found between nephritis in SLE and the presence of AEA reactivity against endolhelial membrane antigens of 38, 41 and 150 kD. These data show that the pattern of AEA reactivity in serum of SLE patients is heterogeneous, and suggest that AEA against a limited number of antigens may be involved in the pathogenesis of nephritis in SLE.
Localization of 20-kD homologous restriction factor (HRF20) in diseased human glomeruli. An immunofluorescence study
Authors: TAMAI H , MATSUO S , FUKATSU A , NISHIKAWA K , SAKAMOTO N , YOSHIOKA K , OKADA N , OKADA H
Published: July 2010

The 20-kD homologous restriction factor (HRF20), which is identical to CD59, is a membrane-associated protein which inhibits the reaction of C9 to form membrane attack complex (MAC) of homologous complements. In various human glomerular diseases deposition of complement components is frequently seen and MAC is reported to associate with immune deposits. Using a specific monoclonal antibody, 1F5, against HRF20, we attempted to study the localization of HRF20 in human glomerulonephritides and to compare the localization of HRF20 with those of immune deposits and MAC. The frozen sections of kidney specimens were fixed in acetone at room temperature before staining. In normal kidneys and kidney specimens from the patients with minimal change nephrotic syndrome, membranous nephropathy. and IgA nephropathy. HRF20 was strongly localized in the peritubular capillaries and along Bowman’s capsules. A weaker but well-defined staining was obtained in the mesangial area and faint staining was seen along the glomerular capillary walls. In contrast, glomerular capillary walls were rather strongly stained in the cases with diffuse lupus nephritis which had subendothelial dense deposits. These data suggest that HRF20 (CD59) is present in the human glomeruli and its expression is enhanced under certain conditions such as lupus nephritis.
Variable gene usage of T cell receptor γ- and δ-chain transcripts expressed in synovia and peripheral blood of patients with rheumatoid arthritis
Authors: OLIVE C , GATENBY P A , SERJEANTSON S W
Published: July 2010

The synovial tissue and fluid of patients with rheumatoid arthritis (RA) contain activated T cells that probably have a central role in the disease process which leads to joint destruction. A subset of T cells, γδ T cells detected at the site of inflammation, may be important in the pathogenesis of the disease. This study investigated variable (V) gene usage of γδ T cell receptors (TcRs) expressed in synovia and peripheral blood of patients with RA by using the polymerase chain reaction (PCR) to amplify TcR γ-and γ-chain transcripts. Most patients showed no restriction in Vγ gene usage since synovial mononuclear cells (SMC) expressed TcR γ-chain transcripts which used the same set of Vγ genes as peripheral blood mononuclear cells (PBMC). In contrast, the majority of patients expressed a restricted SMC Vδ-chain repertoire biased towards Vδ1, but Vδ2 mRNA transcripts were also detected, albeit at low levels in some patients. The TcR δ-chain repertoires of PBMC from healthy control subjects were also characterized. There was variation in the TcR δ-chain repertoires of PBMC from patients when compared with controls, particularly with respect to expression of Vδ4. Vδ4 mRNA transcripts were expressed in PBMC of only two of seven RA patients in contrast with eight of the nine controls (P=0.03). These findings are compatible with reports that γδ T cells in the rheumatoid synovium are reactive to Mycobacterium tuberculosis and that response to M. tuberculosis is restricted to VγL9/Vδ2-bearing T cells, if a superantigen is involved in the pathogenesis of RA.
Flare-up of experimental arthritis in mice with murine recombinant IL-1
Authors: DE VAN LOO A A J , ARNTZ O J , VAN DEN BERG W B
Published: July 2010

Intra-articular injections of murine recombinant IL-1 (mrIL-1) during the chronic phase of antigen-induced arthritis (AIA) induced a flare-up of the smouldering inflammation. The exacerbation was characterized by acute and transient joint swelling and this coincided with the extravascular accumulation of neutrophils. IL-1 injected into arthritic joints of neutropenic mice demonstrated that joint swelling was independent of the neutrophil influx into the joint. Both phenomena were absent when IL-1 was injected into a naive joint. The IL-1-induced flare-up was not T cell mediated as in the antigen-induced flare-up, and suggestive evidence is presented that IL-1 sensitivity depended on the resident macrophage population. This explained why the hypersensitivity is not restricted to the immunologically mediated arthritis but reflects a more general hypersensitivily of previously injured joints, e.g. zymosan-induced arthritis and IL-l-affected joints. In addition, IL-1 could also potentiate the antigen-specific flare-up of chronic AIA and prolongs the duration of the exacerbation. Our data indicate that joints bearing a chronic infiltrate are at risk from exacerbations in two ways: a T cell mediated rechallenge with antigen, and a non-specific reactivation by systemic and local IL-1 generation.
Soluble IL-2 receptor and tumour necrosis factor-α in plasma of haemophilia patients infected with HIV
Authors: NORONHA I L , DANIEL V , SCHIMPF K , OPELZ G
Published: July 2010

We measured plasma concentrations of soluble receptors for IL-2 (sIL-2R) and tumour necrosis factor-a (TNF-a) in 149 haemophilia patients. Soluble IL-2R levels were elevated in 37% of 62 HIV-seroncgative patients (mean 570 ± 27 U/ml versus 361 ± 17 U/ml in the control group, P < 0.0001), in 78% of 68 HIV-seropositive patients (928·49 U/ml, P<0.0001), and in 95% of 19 AIDS/ARC patients (1578 ± 199 U/ml. P<0.0001 compared with controls and with HIV-seronegative patients; P<0.005 compared with HIV-seropositive asymptomatic patients). A negative correlation was observed between sIL-2R, relative and absolute numbers of CD4+ cells (P<0.0001), and CD4/CD8 ratios (P<0.0001). There was also a negative correlation between sIL-2R in plasma and the cellular expression of IL-2R (P<0.001). We found a significant association of sIL-2R and plasma neopterin (P<0.0001). With progression of the disease from HIV-seronegative to seropositive without symptoms and to full manifestation of AIDS/ARC, sIL-2R plasma levels increased. The highest levels were found at the time of diagnosis of AIDS/ARC, but the levels decreased again during the following 18 months. Eight percent of HIV-seronegative patients. 32% of HIV-seropositive patients, and 24% of patients with AIDS/ARC had increased plasma TNF-a. We conclude that sIL-2R and TNF-a plasma levels arc elevated in HIV-infected haemophilia patients and that sIL-2R is a marker for disease progression from asymptomatic HIV-seropositive to AIDS/ARC.
Epstein-Barr virus and HIV play no direct role in persistent generalized lymphadenopathy syndrome
Authors: BOYLE M J , SCULLEY T B , COOPER D A , TURNER J J , PENNY R , SEWELL W A
Published: July 2010

Persistent generalized lymphadenopathy (PGL) and polyclonal B cell activation are features of infection with HIV. Epstein-Barr virus (EBV) and HIV are known to activate B cells in vitro, but whether they are important B cell activators in patients infected with HIV is less clear. In this study, lymph node tissue was obtained from 10 patients with PGL and assessed for evidence of EBV and HIV gene sequences. DNA was extracted and specific viral gene sequences identified using the polymerase chain reaction (PCR). EBV sequences were difficult to detect in the PGL tissue, with a signal intensity similar to that of other benign and malignant lymphoid conditions not associated with EBV. HIV sequences were also rare in the PGL tissue, consistent with HIV infection of the small number of peripheral blood cells and nodal T cells likely to be present in such a sample. These findings suggest that the polyclonal B cell activation typical of HIV is not driven by direct EBV or HIV infection of B cells.
Elevations in cytosolic free Ca2+ are not required to trigger apoptosis in human leukaemia cells
Authors: LENNON S V , KILFEATHER S A , HALLETT M B , CAMPBELL A K , COTTER T G
Published: July 2010

Previous studies have indicated that Ca2+ is a trigger for apoptosis (programmed cell death) in thymocytes and related cell lines. Recently we have shown that levels of apoptosis in leukaemic cells are diminished in Ca2+-deficient conditions, indicating that Ca2+ may be important in the mechanism of apoptosis in these cells. In the present study we investigated the possibility that Ca2+ serves as a trigger for apoptosis in the human leukaemic cell line, HL-60. Using fura-2 to measure cytosolic free Ca2+ concentrations, [Ca2+]i, in cell suspensions, and by using ratio imaging of fura-2 in single cells, we did not observe an early significant increase in [Ca2+]i, in HL-60 cells undergoing apoptosis. The latter stages of apoptosis were, however, accompanied by increasing [Ca2+]i, these increases were apparently a result of, rather than a cause of, apoptosis. Furthermore, apoptosis could be induced in HL-60 cells under conditions of vastly reduced [Ca2+]i achieved by loading these cells with fura-2 in the presence of EGTA. These results indicate that elevation of [Ca2+]i is not a prerequisite for apoptosis in HL-60 cells and that apoptosis can occur in these cells in the presence of low[Ca2+]i.
Cytomegalovirus (CMV) infection, CD4+ lymphocyte counts and the development of AIDS in HIV-1-infected haemophiliac patients
Authors: WEBSTER A , PHILLIPS A N , LEE C A , JANOSSY G , KERNOFF P B , GRIFFITHS P D
Published: July 2010

After a maximum of 11 years (median 8·3 years) from the time of HIV seroconversion. 25 out of 59 (42%) of CMV-seropositive haemophiliacs had progressed to AIDS, as opposed to eight out of 50 (16%) CMV scroncgatives. The age-adjusted relative risk for AIDS among CMV scropositives was 2·4 (P= 003). In order to determine how this adverse effect is mediated, the mean rate of decline in serial CD4+ lymphocyte counts was studied. CD4+ lymphocyte counts tended to decline more rapidly in CMV seropositives than in seronegatives (−0·087 ± 109/l per annum versus-0·082 ± 109/l per annum), but this difference did not reach statistical significance. The average CD4+ lymphocyte count at the lime of HIV seroconversion was estimated to be similar in CMV seropositives and negatives, because in HIV-l-ncgative haemophiliacs the CD4+ counts were virtually identical, after adjustment for age (0·94 ± 10+/l and 0·97 ± 109/l. respectively). The median CD4+ cell count at which AIDS developed was higher in the CMV-seropositive group (0·07 ± 109/l) than in the seronegative group (0·04 ± 109/l). but this difference did not reach statistical significance. We conclude from these findings that the adverse effect of CMV is not wholly mediated via a more rapid loss of CD4+ cells. We discuss other processes that may be mediated by CMV, such as a functional deficiency of residual CD4+ cells, or dissemination of HIV in other organs, which may be important in determining the earlier onset of AIDS among CMV-seropositive subjects.
LAM-1/Leu 8 antigen is expressed on portal, but not on tabular intrahepatic mononuclear cells in inflammatory liver disease
Authors: VOLPES R , VAN DEN OORD J J , DESMET V J
Published: July 2010

The expression of the selectin receptor LAM-1/Lcu 8 was analysed in normal and in inflamed liver tissue, and its expression on mononuclear inflammatory cells was correlated with their topographical distribution in various compartments of the inflamed liver, in order to obtain new insights on possible molecular mechanisms involved in the traffic of mononuclear inflammatory cells throughout the diseased hepatic parenchyma. In normal liver tissue, few scattered mononuclear cells in portal and lobular parenchyma corresponded to both CD4+ and CD8+, as well as to CD45RA+ (2H4+) naive and CD45RO+ (UCHL1 +) memory T cells, and were LAM-1 /Leu 8+. In acute and chronic inflamed liver biopsies. CD45RO+ (UCHL1 +)CD4+ and CD8+ memory T cells largely predominated in both portal and lobular parenchyma. The expression of LAM- 1/Leu 8 antigen on these memory T cells varied according to their localization in the liver parenchyma, and it was not correlated with specific aetiological causes. In acute hepatitis, the vast majority of T lymphocytes were LAM-1/Leu 8. In chronic active hepatitis, memory T cells in portal tracts expressed LAM-1 /Leu 8. whereas virtually all intralobular T cells accumulating in areas of periportal and intralobular inflammation were LAM-1/ Leu 8. In chronic persistent hepatitis, the LAM-1/Leu 8+ T cells largely predominated among the numerous mononuclear inflammatory cells within enlarged portal tracts, whereas LAM-1/Leu 8 T cells were restricted to areas of intralobular ‘spotty’ inflammation. Therefore, two phenotypical populations can be recognized among the memory T cells in inflamed liver tissue, according to their topographical localization: LAM-1/Leu 8+ Tcells predominating in portal tracts, and LAM-1/Leu 8 T cells predominating in the lobular parenchyma. These data show that during their migration through the inflamed liver parenchyma, memory T lymphocytes undergo phenotyical changes (LAM-1/Leu 8 shedding) according to their localization in different liver compartments (portal tracts vs. lobular parenchyma), suggesting multiple cellular and molecular mechanisms involved in the regulation of the leucocyte traffic through inflamed liver tissue.
Repertoire of CDS+ and CD5− cord blood B cells: specificity and expression of VH I and VH III associated idiotopes
Authors: MACKENZIE L E , MAGEED R A , YOUINOU P Y , YUKSEL B , JEFFERIS R , LYDYARD P M
Published: July 2010

Epstein-Barr (EBV)-immorlalized B cell clones were established from CD5+ and CD5 cord blood B cells separated by flow cytometry. We have previously shown that lgM from many of the clones was polyreacti ve, exhibiting reactivity with a number of autoantigens. In this study, IgM produced by the clones was analysed by MoAb for the expression of cross-reactive idiotypes (CRI) associated with rheumatoid factor paraproteins and from defined VH and Vk subgroups of immunoglobulin heavy and light chains. IgM produced by clones established from CD5+ and CDS B cells expressed the VH I associated idiotope G8. Furthermore, IgM produced by both sets of clones exhibited a similar frequency of VH III heavy chain subgroup expression, as determined by reactivity with staphylo-coccal protein A (SpA) and VH III-associated CRI expression (B6 and/or DI2). In contrast, expression of the V, III-associated 17.109 CRI was significantly higher in IgM antibodies produced by clones established from CD5+ compared with the CD5– clones(32 versus 5%: P < 0·05). Analysis of the VH and VL subgroup expression by IgM produced by the CD5’ and CD5 cord blood clones, and their autoantigen reactivity profile did not reveal restriction or selection within CD5+ and CD5–populations. However, our data suggest that differences may exist in the expression of certain germline genes between CD5+ and CD5– cord blood B cells and might indicate an expansion of CD5+ B cells within the fetal environment.
Modulation of human T cell functions by surface sulphydryl groups: differential effects on IL-2 production and responsiveness
Authors: SMITH S , BROWN-GALATOLA C H , HALL N D
Published: July 2010

An impermeable thiol blocker has been used to investigate the role of sulphydryl (SH) groups in the production of and responsiveness to IL-2 by normal human T lymphocytes. Surface SH blockade of mononuclear cells prior to incubation with mitogen (phytohaemagglutinin, concanavalin A, CD3 MoAb) had no effect on production of IL-2 but markedly impaired cellular responsiveness to exogenous IL-2. Studies using MoAbs indicated that this effect was accompanied by decreased expression of both the CD25 and p75 subunits of the IL-2 receptor. Blocking surface SH groups did not affect binding of IL-2 to p75 on unstimulated mononuclcar cells, but inhibited binding to high-affinity receptors on a T lymphoma cell line. The data are consistent with the hypothesis that sulphydryl groups on the IL-2 receptor are required for its function and may be involved in the interaction of the CD25 and p75 subunits leading to generation of the high-affinity binding site. The surface thiol identified on the IL-2 receptor may be a candidate for oxidation on cells from patients with chronic inflammatory diseases such as rheumatoid arthritis and thus contribute to the aberrant function of T cells in these patients.
Localization of tumour necrosis factor production in cells at the materno/fetal interface in human pregnancy
Authors: VINCE G , SHORTER S , STARKEY P , HUMPHREYS J , CLOVER L , WILKINS T , SARGENT I , REDMAN C
Published: July 2010

Biologically active tumour necrosis factor (TNF) was detected in medium conditioned by incubation with cxplants of human pregnancy decidua or fetal chorionic villous tissue, taken in the first trimester and at term. Addition of endotoxin increased TNF release in most cases. ELISA assays gave similar results for TNF–α and also demonstrated low levels of TNF–β. Using cell populations purified by flow cytometry, secretion of biologically active TNF was shown to be localized to the macrophages. Cytotrophoblast purified from term amniochorion produced no TNF. Both decidual and chorionic villous tissue at term contained mRNA for TNF–α and TNF–β. TNF–α mRNA was confined to decidual macrophages in first trimester tissue, and was not present in chorionic cytolrophoblast. TNF–β mRNA, in contrast, was detected in both macrophage and no-acrophage populations in term dccidua.
Characterization of IL-1 inhibitory factor released from human alveolar macrophages as IL-1 receptor antagonist
Authors: TAKEUCHI M , NAGAI S , NAKADA H , AUNG H , SATAKE N , KANESHIMA H , IZUMI T
Published: July 2010

IL-1 possesses pleiotropic properties on various cells and its activity may be stringently regulated in several ways. We have previously reported that both IL-1 and its inhibitory factor are concomitantly released from alveolar macrophages in both healthy subjects and patients with chronic inflammatory lung diseases. An increase in IL-1 activities and a decrease in inhibitory activities arc characteristics found in both healthy smokers and patients with interstitial lung diseases. In this study, we further examined the biological properties of IL-1 inhibitory factor. The inhibitor exhibited a dose-dependent specific inhibition of an augmentation by IL-1 of PH-nduced murinc thymocyte proliferation, while no inhibition of the augmentation by IL-2, IL-4, IL-6, or tumour necrosis factor (TNF) was found. 125I-labelled IL-1α binding on PHA-stimulated murinc thymocytes revealed two types of IL-1 binding sites. 44 sites/cell with a Kd of 2.7 ± 10−10 m and 230 sites/cell with a Kd of 2.5 ± 10−9 m. Alveolar macrophage culture supernatants blocked the binding of labelled IL-1 to the IL-1 receptor in a dos-ependent fashion. Scatchard plot analysis revealed that the inhibitory factor in the supernatants blocked the binding competitively. These results indicate that alveolar macrophages produce a specific IL-1 inhibitory factor, functioning as an IL-1 receptor antagonist.
During HIV-1 infection most blood dendritic cells are not productively infected and can induce allogeneic CD4+ T cells clonal expansion
Authors: CAMERON P U , FORSUM U , TEPPLER H , GRANELLI-PIPERNO A , STEINMAN R M
Published: July 2010

We have considered the possibility that antigen-presenting cells of the dendritic cell lineage may be infected in vivo and spread HIV-1 at the time dendritic cells initiate the clonal expansion of antigen-specific T cells. Dendritic cells were isolated from 25 HIV-1–infected subjects (CDC stages II- IV). Fewer dendritic cells were recovered from most infected subjects. Reduced numbers of total non-T cells were also found in these patients, so that preferential loss of dendritic cells did not occur. Dendritic cell function was assessed by stimulatory capacity for allogeneic CD4+ T cells in the mixed leucocyte reaction (MLR). Potent M LR stimulator activity was retained in the dendritic cell-enriched populations from HIV-infected patients. Seven out of nine patients without AIDS (asymptomatic, lymphadenopathy or ARC) and three out of six patients with AIDS had proliferative responses equivalent to those induced by dendritic cells from controls. Dendritic cells from HIV+ subjects were able to initiate the expansion of allogeneic CD4+ T cell clones with cloning efficiency not different from controls and without evidence of cytopathic effect in the expanding CD4+ clones. In situ hybridization of the different mononuclear cell populations with a gag-specific riboprobe demonstrated positive cells in the T cell fractions of 12 of the 15 patients tested. None of the asymptomatic or ARC patients had riboprobe-positive cells in the dendritic cell-enriched populations. Four out of nine patients with AIDS had cells positive for HIV-1 expression in the dendritic cell-enriched fraction. However, the positive cells had the nuclear profile of lymphocytes, and by cytofluorography some residual low-density T cells were present. By limiting dilution and polymerase chain reaction (PCR), CD4+ lymphocytes carried HIV provirus in inocula of 500–5000 cells, while provirus could only be detected in 50000 cells from the dendritic cell-enriched fraction. The latter signal may be due to the demonstrated levels of T cell contamination. Our data indicate that productive or latent HIV-1 infection of blood dendritic cells in vivo is rare, certainly no greater than in T lymphocytes, and that in vitro dendritic cell preparations from patients can expand CD4’ T cells efficiently and therefore may be able to expand T cells with immunotherapeutic activity.
Laboratory control values for CD4 and CD8 T lymphocytes. Implications for HIV-1 diagnosis
Authors: BOFILL M , JANOSSY G , LEE C A , MACDONALD-BURNS D , PHILLIPS A N , SABIN C , TIMMS A , JOHNSON M A , KERNOFF P B A
Published: July 2010

With the advent of standard flow cytometric methods using two-colour fluorescence on samples of whole blood, it is possible to establish the ranges of CD3. CD 4 and CDS T lymphocyte subsets in the routine laboratory, and also to assist the definition of HIV-1-related deviations from these normal values. In 676 HIV-1-seronegative individuals the lymphocyte subset percentages and absolute counts were determined. The samples taken mostly in the morning. The groups included heterosexual controls, people with various clotting disorders but without lymphocyte abnormalities as well as seronegative homosexual men as the appropriate controls for the HIV-1-infected groups. The stability of CD4% and CD8% values was demonstrated throughout life, and in children CD4 values < 25% could be regarded as abnormal. The absolute counts of all T cell subsets decreased from birth until the age of 10 years. In adolescents and adults the absolute numbers (mean±s.d.) of lymphocytes, CD3, CD4 and CD8 cells were 1·90±0·55, 1·45±0·46, 0·83±0·29 and 0·56± 0·23 ± 109/l, respectively. In patients with haemophilia A and B the mean values did not differ significantly. In homosexual men higher CD8 levels were seen compared with heterosexual men and 27% had an inverted CD4/CD8 ratio but mostly without CD4 lymphopenia (CD4<0·4 ± 109/l). However, some healthy uninfected people were‘physiologically’ lymphopenic without having inverted CD4/CD8 ratios. When the variations‘within persons’ were studied longitudinally over a 5-year period, the absolute CD4 counts tended to be fixed at different levels. As a marked contrast, over 60% of asymptomatic HIV-1+ patients exhibited low CD4 counts <0·4 ± 109/l together with inverted CD4/CD8 ratios. Such combined changes among the heterosexual and HIV-1-seronegative homosexual groups were as rare as 1·4% and 3%, respectively. For this reason, when the lymphocyte tests show <0·4 ± 109/l CD4 count and a CD4/CD8 ratio of less than unity, the individuals need to be investigated further for chronicity of this disorder, the signs of viral infections such as HIV-1 and other causes of immunodeficiency.
Tumour necrosis factor, IL-1 and IL-6 in bronchoalveolar washings and theirin vitroproduction duringNippostrongylus brasiliensisinfection
Authors: BENBERNOU N , MATSIOTA-BERNARD P , JOLIVET C , OUGEN P , GUENOUNOU M
Published: July 2010

Bronchoalveolar washings (BAW) were obtained from rats primarily infected with N. brasiliensis during the early infection stage that coincides with the lung passage of the parasite and the recruitment of inflammatory cells. BAW were tested for IL-1, IL-6 and tumour necrosis factor (TNF) activities. We found that IL-1 production occurred only on day 1 post infection and ceased thereafter. IL-6 activity was present as from day 1 with a maximum on day 3 post infection and then returned to its normal levels on day 5 post infection. TNF activity was not recovered in BAW at any time of the early infection. Results obtained from the in vitro culture of BAW-adhcrent cells demonstrated that on day I post infection IL-1, but also large amounts of TNF were produced spontaneously, whereas IL-6 was continuously released. Bacterial lipopolysaccharide (LPS) stimulation of the cell culture resulted in an amplification of the cytokine production. Our results suggest that pulmonary cytokincs detected in BAW were at least in part produced by alveolar macrophages. Furthermore, the kinetics of IL-1, TNF and IL-6 production show that these monokines are induced at different times during the course of infection, suggesting that cytokine production may follow different regulation patterns during the early phase of N. bransiliensis infection.
Apoptosis in human thymocytes after treatment with glucocorticoids
Authors: NIETO M A , GONZÁLEZ A , GAMBÓN F , DÍAZ-ESPADA F , LÓPEZ-RIVAS A
Published: July 2010

Treatment of unfractionated human thymocytes in culture with the synthetic glucocorticoid dexamethasone induced cell death, as measured by trypan blue exclusion, after several hours of incubation. In purified subsets of human cortical and medullary thymocytes dexamethasone caused cell lysis with similar kinetics in both populations; 50% of thymocytes were killed after 20–24 h of incubation with the steroid. The mechanism of dexamethasone-induced cell death seems to correspond to apoptosis since degradation of DNA into oligonucleosome-sized fragments could be observed in the cultures treated with the steroid. A certain degree of DNA fragmentation and cell death could also be observed in control cultures of thymocytes. In contrast, peripheral T lymphocytes were resistant to the cytolytic effect of glucocorticoid hormone. The killing of human thymocytes by dexamethasone was inhibited by cycloheximide, suggesting that this cell death program requires a fully operating protein synthesis machinery and perhaps the induction of new proteins.
Relationship between a novel human cytotoxin (factor 2) produced by a B cell line (Karpas 160) and phorbol-myristateacetate-associated cytotoxicity
Authors: NI J , KARPAS A
Published: July 2010

We found that 160b cells, a subclone of Karpas 160 human B cell line, spontaneously secreted a novel cytotoxin, factor 2 (F2). F2 was also extracted from the cells by 60% ammonium sulphate. 0·5% CHAPS and 0·28% Triton X-114. We were able to show that phorbol myristate acetate (PMA) greatly enhanced the production of F2, and PMA may also account for part of the putative F2 cytotoxic activity to K562 cells in crude preparations. We compared the cytotoxic effect of F2 with PMA-associated F2-like cytotoxicity to K562 cells as well as the adequacy of our schemes to purified F2 with regard to its separation from PMA. We found that it was possible to separate PMA from F2 preparations by gel filtration and Rotofor preparative isoelectric focusing. The fate of PMA was also monitored with 3H-PMA and chromatographic profiles of 3H-PMA were studied using DE52 and gel filtration chromatography. We were able to establish that less than 2·9% of the cytotoxicity to K562 was due to PMA. We also found that the radioactive peaks and cytotoxicity peaks to K562 were not well correlated, indicating that the cytotoxicity was not mainly due to remaining PMA. Activated charcoal removed virtually all F2 and PMA but not tumour necrosis factor activity. Our results also showed that cytotoxicity to K562 resulting from F2 or PMA-associated proteins had different physicochemical properties, indicating that they are different molecular entities. These findings are consistent with the earlier observation that 160 cells produce F2 spontaneously and that PMA can amplify its production significantly.
Characterization of the lymphocytic alveolitis in visna-maedi virus-induced interstitial lung disease of sheep
Authors: CORDIER G , GUIGUEN F , CADORÉ J L , COZON G , JACQUIER M F , MORNEX J F
Published: July 2010

In order to investigate the contribution of lymphocytes to interstitial lung disease in animals with visna-maedi infection, we studied in parallel bronchoalveolar cells and lung tissue from slaughterhouse animals (n = 29) and from colostrum-deprived lambs transtracheally inoculated with field isolates of visna-maedi virus (n = 9) or saline (n = 6). Lymphocyte subpopulations were identified in bronchoalveolar lavagc by immunofluorescence and flow cylomctry analysis and in lung tissue using indirect immunohistochemistry. In infected animals a lymphocytic alveolitis containing CD4 and CD8 lymphocytes was observed. Pcribronchovascular lymphoid nodules comprise mostly CD4 lymphocytes. Alveolar lymphocytes of both subsets displayed increased expression of MHC class II antigens in animals with naturally occurring maedi but not in experimentally infected ones. A sequential process of lymphocyte attraction and activation is likely to occur in vivo as part of the alveolitis.
Alterations in mononuclear cell tumour necrosis factor-alpha (TNF-a) response in patients on long term cuprophane haemodialysis
Authors: ANNENKOV A Y , STROKOV A G , BARANOVA F S
Published: July 2010

We have investigated TNF-a secretory response of peripheral blood mononuclear cells (PBMC) from 13 uraemic patients undergoing regular haemodialysis with cuprophane membrane (CM). Sixteen healthy subjects and five uraemic patients under conservative therapy were also studied as controls. Cells of haemodialysis patients exhibited increased TNF-a release in vitro in the absence of activating stimuli other than culture conditions, as compared with normal and uraemic controls. In contrast to normal cells, this spontaneous secretion of TNF-a from dialysis PBMC could not be significantly reduced by addition of polymyxin B to culture medium, thus indicating its independence of trace amount of lipopolysaccharide (LPS) present in the medium as contaminant. Furthermore, prc-dialysis PBMC were considerably more sensitive to stimulation with It)7 pg/ml of LPS under in vitro culture conditions than normal and uraemic controls. To elucidate a role of direct contact with CM in stimulation of TNF-a release from monocytes. PBMC were cultured on CM in vitro. Contact with CM stimulated TNF-a secretion from PBMC above the level of cells cultured on tissue culture plastic. This response persisted with time in culture in contrast to a transient LPS-induccd TNF-a release. Furthermore. PBMC stimulated by contact with CM for 2 days did not lose the capacity to secrete TNF-a in response to a subsequent LPS stimululion, while a 2-day treatment of cells with LPS was followed by LPS refractory slate. Therefore, direct contact with CM induces in PBMC a long-lasting TNF-a response which is not down-regulated by the acquisition of refractoriness in a manner similar to that which occurs in the case of LPS stimulation. These in vitro findings provide a possible explanation of the observation that predialysis PBMC exhibit elevated TNF-a secretory capacity.
The role of O-antigen polysaccharide in the activation of neutrophils by lipopolysaccharides ofSalmonellaspecies
Authors: RASOOL O , NNALUE N A , JARSTRAND C
Published: July 2010

Activation of neutrophils by lipid A, O-antigen polysaccharides (PS) and smooth lipopolysacchar ides (LPS) isolated from Salmonella choleraesuis (O-6,7) and Salmonella typhimurium (O-4,5,12) was investigated. The methods used were assays for lysozyme release and for nitroblue tetrazolium (NBT) reduction which measures the level of oxidative metabolism of neutrophils. LPS from both species stimulated neutrophils to the same extent in the presence of autologous plasma. In the absence of plasma only the O-6,7 LPS activated neutrophils. Lipid A of PS isolated from both LPS either did not activate neutrophils of did so only at very high concentrations when tested in the presence of plasma; in the absence of plasma no activation occurred. The data indicate that both PS and lipid A segments of LPS are required for activation of neutrophils by LPS. We also deduce that plasma, probably complement, is required for the interaction of some LPS, e.g. 0–4,5,12 with neutrophils whereas other LPS, e.g. 0–6,7 can interact directly and activate neutrophils.
Endothelial cell damage in primary biliary cirrhosis: influence of cholestasis and immunological mechanisms
Authors: BLANN A D , BABBS C , NEUBERGER J M
Published: July 2010

In a study looking for evidence of endothelial cell damage in primary biliary cirrhosis, serum from patients was found to be significantly more cytotoxic to cultured endothelial cells in vitro than normal control serum (P<0·001). Serum also contained higher levels of von Willebrand factor antigen (vWFAg, a specific product of the endothelium) than control serum (P<0·001). Cytotoxicity correlated with serum levels of vWFAg (P<0·05) and with serum bilirubin (P<0·05). No correlations were found between cytotoxicity and circulating immune complexes. C-reaetive protein or CH50. The study was controlled by serum from patients with other liver diseases. In these samples vWFAg (P<0·003), and bilirubin levels (P<0·001) were also raised relative to normal controls. vWFAg levels again correlated with levels of bilirubin (P<0·05). Tissue culture experiments showed that purified bilirubin was cytotoxic to human umbilical vein endothelial cells and induced the release of vWFAg. The case report of a patient with obstructive jaundice again indicated that levels of bilirubin and vWFAg were related. These results suggest that endothelial cell damage occurs in both primary biliary cirrhosis and to a lesser extent in other liver diseases, and may be mediated by cholestasis, not by humoral immunological mechanisms.
Endoglin: a 180-kD endothelial cell and macrophage restricted differentiation molecule
Authors: O'CONNELL P J , McKENZIE A , FISICARO N , ROCKMAN S P , PEARSE M J , d'APICE A J F
Published: July 2010

A MoAb RMAC8 was generated by immunizing Balb/c mice with cultured human umbilical vein endothelialcells (HUVE). The molecule recognized had a mol. wt of 180 kD non-reduced, 95 kD after reduction and 66 kD in its reduced and n-dcglycosylated form. Sequential immunoprecipilalion studies with the MoAb 44G4, which recognizes the o- and n-glycosylated homodimer endoglin, showed that both MoAbs recognize the same molecule on HUVE and phorbol myristate acetate (PMA)-stimulated U937 cells. The distribution of the RMAC8-recognized molecule was the same as that described for endoglin. i.e. arterial and venous endothelium, myelomonocytic and pre-B leukaemia cells and cell lines; however, unlike 44G4, RMAC8 also reacted weakly with monocytcs and strongly with in vitro differentiated macrophages as well as peritoneal and alveolar macrophages. This distribution of endoglin was confirmed by Northern blot analysis using a full length endoglin cDNA probe. These studies suggest that endoglin is a differentiation marker on macrophages.
A hereditary immunodeficiency characterized by CD8+ T lymphocyte deficiency and impaired lymphocyte activation
Authors: MONAFO W J , POLMAR S H , NEUDORF S , MATHER A , FILIPOVICH A H
Published: July 2010

An unusual form of severe combined immunodeficiency in children from two different families was associated with absence of CD8+ T lymphocytes and normal numbers of CD4+ T lymphocytes that did not respond to stimulation by non-specific mitogens, specific antibodies against the T cell receptor or specific antigens. The defect in the CD4+ cells was bypassed by activating agents which are independent of the T cell receptor. The combination of an activation defect and selective depletion of CD8+ T lymphocytes suggests that the defective pathway is important in the differentiation of immature thymocytes as well as the proliferation of mature lymphocytes.
Induction of adjuvant arthritis in mice
Authors: KNIGHT B , KATZ D R , ISENBERG D A , IBRAHIM M A , Le PAGE S , HUTCHINGS P , SCHWARTZ R S , COOKE A
Published: July 2010

Adjuvant arthritis, induced by injections of Freund's complete adjuvant into the footpads of some rat strains, has been recognized as a useful animal model for many years. There has, however, been notable lack of success in reproducing this model in other species. We now describe the development of adjuvant arthritis in healthy strain mice approximately 2 months after injection of Freund's complete adjuvant. Although the clinical appearance of the mice and the joint histopathology closely resemble the adjuvant arthritis reported in the rat. we were unable to detect rheumatoid factor in sera from the affected animals. In parallel studies of T cell proliferation, affected animals responded to some mycobaclcrial antigens but not to the 65-kD heat shock protein of Mycobacterium tuberculosis, suggesting that some other epitope is important in the development of the disease.
Activated T-lymphocytes induce growth inhibition and prostaglandin E2 release from syngeneic glomerular mesangial cells
Authors: RADEKE H H , SCHWINZER B , RESCH K
Published: July 2010

The concept of an active role of T lymphocytes in the initiation and development of autoimmune glomerulonephritis has gradually evolved from recent investigations. In the present study we started in a murinc coculture system to directly examine cellular interactions of intrinsic glomerular mesangial cells (MC) and syngeneic T lymphocytes. Lymph node lymphocytes and. moreover, cloned T helper cells specifically affected syngeneic proliferating MC. causing growth inhibition and prostaglandin E2 (PGE2) release. The T cell specificity of mesangial cell responses was confirmed by demonstrating (i) that MC coculturcd with other cell types showed no reaction and (ii) that additional activation of T lymphocytes by IL-2 or concanavalin A significantly enhanced the MC responses. Subsequently, we confirmed the presence of T cell factors in the supernatants responsible for the observed effects: interferon-gamma (IFN-γ) and tumour necrosis factor (TNF). Experiments with combinations of recombinant mouse IFN-γ and human lymphotoxin or TNF-α showed that these lymphokines could substitute for the direct T lymphocyte effects causing a synergistic growth inhibition and PGE2 release from mouse MC. The observed lymphokine activities were not due to mesangiolysis as shown by neutral red uptake of MC. Pointing to the essential role of T helper cell-specific products, IFN-γ antibodies abolished both the IFN-γ and the combined IFN-γ/TNF-α effect. Thus, our investigations with syngeneic MC-lymphocyte cocultures demonstrated that cultured MC specifically responded to T lymphocytes and their products.
Antibodies to Mycobacterium paratuberculosis-specific protein antigens in Crohn's disease
Authors: ELSAGHIER A , PRANTERA C , MORENO C , IVANYI J
Published: July 2010

The possible role of infection with Mycobacterium paratuberculosis (MAP) for the etiopathogenesis of Crohn's disease (CD) has been a matter of long-term controversy. In addition to similarities with the pathology of ruminant paratuberculosis, DNA fingerprinting confirmed the organism isolated from gut tissue, but the specificity of the immune repertoire has not as yet been evaluated. We report here on a serological study of 29 patients with CD, 20 patients with ulcerative colitis and 18 healthy control subjects, using three antigens attributed with species-specificity and selective immunogenicity following MAP infection. Antibodies binding to the 38-kD band of MAP extract were demonstrable by the Western blot technique in 57% of CD patients. Antibody levels to the 24-kD (p24BCD) cathodic bands, determined by competition ELISA using a monospecific murine antiserum, and to the 18-kD protease-resistant purified bacterioferritin, delected by standard ELISA, were significantly elevated in 53% of CD patients. However, these three antibody specificities tested in individual CD patients did not show any correlation with each other. Thus, 18% of patients were positive for all three specificities, whilst 84% had antibodies to at least one of the specific antigens. Although the exact proportion of affected patients is yet to be defined, the serological results obtained support the view that MAP infection may play an ctiological role in Crohn's disease.
Expression and gene transcript of Fc receptors for IgG, HLA class II antigens and Langerhans cells in human cervico-vaginal epithelium
Authors: HUSSAIN L A , KELLY C G , FELLOWES R , HECHT E-M , WILSON J , CHAPMAN M , LEHNER T
Published: July 2010

The mechanism of transmission of HIV from the male to the female genital tract or in the reverse order is not clear. CD4 glycoprotein is the receptor for HIV and Langerhans cells and the related dendritic cells could play a role in the initial transmission of HIV. Fc receptors (FcR) for IgG might be involved in antibody-mediated binding of HIV. We carried out an immunohistological study of normal human cervical and vaginal epithelia for the presence of CD4 glycoprotein, Langerhans cells and FcR to IgG. CD4+ glycoprotein was not found in the vaginal or cervical epithelium, with the exception of a few endocervical epithelial cells. A small number of CD4+ mononuclear cells were found in the endocervical epithelium of a third of the specimens but a large number of CD4+ cells was found in the submucosa of most of the cervical and vaginal specimens. Langerhans cells expressing CD4, HLA class II, FcγR2 and FcγR3 were detected in most vaginal, ectocervical and transformation zone epithelia and in 9/14 endocervical tissues. FcγR3 was detected in about two-thirds of the columnar endocervical epithelium and the transformation zone. A smaller number of specimens expressed FcγR2 in these epithelia, but FcγRl was not detected. We then demonstrated mRNA for FcγR3 in the columnar endocervical epithelial cells and transformation zone by in situ hybridization, usinga CD16-RNA probe. FeγR3 and FcγR2 gene transcripts were also found in fetal cervical tissue by applying the polymerase chain reaction to amplify portions of the FcγR3 and FcγR2 coding sequences in cDNA prepared from fetal RNA. HLA-DR was found in the endocervical cells, transformation zone and in Langerhans cells of all specimens. The presence of Langerhans cells, Fcγ receptors and HLA class II antigen offers three potential mechanisms for cervico-vaginal HIV transmission: (i) direct HIV infection of Langerhans cells, (ii) binding of HIV antibody complexes to cervical epithelial Fcγ receptors and (iii) binding of HIV infected CD4+ cells to cervical HLA class II antigen which may infect these or the adjacent CD4+ cells.