bluuebunny/crossref_metadata_2025 huggingface dataset:
Substance P-induced granulocyte infiltration in mouse skin: the mast cell-dependent granulocyte infiltration by the N-terminal peptide is enhanced by the activation of vascular endothelial cells by the C-terminal peptide
Authors: IWAMOTO I , TOMOE S , TOMIOKA H , YOSHIDA S
Published: July 2010

Previous studies have shown that substance P induces granulocyte infiltration in mouse skin, which is mediated through mast cell degranulation. However, it is not yet known whether the direct effect of substance P on vascular endothclial cells is involved in the granulocyte infiltration in the skin. To solve this issue, we used the N-terminal peptide substance P1–9 (SP1–9), which is active for mast cells but inactive for vascular endolhelial cells, and the c-terminal peptide SP6–11, which is active for vascular endothelial cells but inactive for mast cells, since substance P activates both mast cells and vascular endothelial cells. The subcutaneous administration of substance P (10−7−10−5 m) caused granulocyte (neutrophil and eosinophil) infiltration in the skin of BALB/c mice 6 h after the injection. SP1–9 (10−5–10−4 m) also caused granulocyte infiltration of mouse skin which was associated with mast cell degranulation. In contrast. SP6–11 (10−7-10−4 M), which was found to increase the vascular permeability of endothelial cells in mouse skin, induced no significant granulocyte infiltration nor mast cell degranulation. However, SP6–11 (10−5 10−4 m) enhanced SP1–9 -induced granulocyte infiltration in the skin without any significant increase in mast cell degranulation. We conclude that substance P causes granulocyte infiltration in mouse skin through both mast cell degranulation induced by the N-lerminal peptide of substance P and the activation of vascular endothelial cells induced by the c-terminal peptide of substance P.
Quantitative studies of monoclonal antibody 5-1-6-induced proteinuric state in rats
Authors: KAWACHI H , MATSUI K , ORIKASA M , MORIOKA T , OITE T , SHIMIZU F
Published: July 2010

Murine monoclonal antibody (MoAb) 5-1-6 was already reported to bind to epithelial cell fool processes and to cause proteinuria in rats. In vivo kinetics of the injected MoAb 5-1-6, relationship between the quantity of kidney-binding antibody and proteinuria. and changes in the amount of antigenic molecule recognized by this MoAb in the proteinuric state were studied. The amount of total kidney-binding antibody (TK Ab) as determined I h after a 2 mg administration was 50.8 ± 10.4 μg/2 kidneys, and TKAb declined to 1.9 ± 0.4 at day 15. The minimum dose of MoAb required to induce proteinuria was 125 μg as the injected dose. This dose corresponded to 12-8 μg of TKAb at I h and 0.34 μg of TKAb at day 5. The amount of MoAb 5-1 -6 binding to isolated normal glomeruli was also shown to exceed 147-7 μg/76000 glomeruli, indicating proteinuria to be induced provided more than 8.7% (= 12.8/147.7) of the critical epitopes is specifically occupied by MoAb. The total amount of MoAb 5-1-6 bound to glomeruli in vivo and in vitro was assayed with [125I]-labelled anti-mouse IgG. The amount of [125I] anti-mouse IgG bound to glomeruli was 6.93 ± 0.45 μg/10 000 glomeruli from rat 5 days after this MoAb injection and 26.58 ± 0.66 μg/10000 control glomeruli, indicating the decrease in the number of MoAb 5-1-6-rccognizcd antigen molecules in glomeruli isolated from the rat in proteinuric state induced by this MoAb. Thus, the MoAb 5-1-6-recognized molecule itself may principally function to regulate the permeability of the glomerular capillary wall and the decrease of the molecule may lead to proteinuria.
Levels of soluble IL-2 receptor in plasma from asthmatics. Correlations with blood eosinophilia, lung function, and corticosteroid therapy
Authors: LASSALLE P , SERGANT M , DELNESTE Y , COSSET P , WALLAERT B , ZANDECKI M , CAPRON A , JOSEPH M , TONNEL A B
Published: July 2010

Evidence now suggests that cosinophils and T lymphocytes infiltrating bronchial tissues may play a key role in the pathophysiology of asthma. Circulating cosinophils, lung function, and plasma soluble IL-2 receptor (sIL-2R) were measured in 42 asthmatic patients referred for symptomatic asthma. The patients were divided into two groups based on the presence or absence of atopy. The group of non-atopic asthmatics was further divided according to the patients’ requirement for long term oral corticosteroids. The mean sIL-2R ± s.d. was 36.3 ± 9.9 pM in the control group, 28.9 ± 9.2 pM in the atopic asthmatics, 43.3 ±18.07 pM in the non-atopic asthmatics without oral steroid therapy, but was increased in the steroid-treated group (62.2 ±19-3 pM, P<0.01). A significant correlation was found between I-FEV1 and circulating cosinophils in atopic asthmatics and in non-atopic asthmatics without oral corticosteroid therapy, but not in the steroid-treated group. Furthermore, significant correlations were found between sIL-2R and FEV1 and between sIL-2R and blood eosinophils, in the group of non-atopic asthmatics not on oral steroid therapy. No such correlations were evidenced in the other groups of asthmatics. Similar results were obtained during the clinical course of three non-atopic patients followed for more than I year. These data suggest that T cell activation appears more prominent in non-atopic asthma than in atopic asthma. Moreover, it appears that T cell activation can occur in severe forms of asthma despite steroid treatment. Finally, the results suggest a possible link between T cell activation, eosinophils. and lung function, which may reflect a particular pathogenetic mechanism involved in non-atopic asthma.
Enhancement of the antigen-presenting function of monocytes by cholesterol: possible relevance to inflammatory mechanisms in extrinsic allergic alveolitis and atherosclerosis
Authors: HUGHES D A , TOWNSEND P J , HASLAM P L
Published: July 2010

Extrinsic allergic alveolitis (EAA) (synonym: hypersensitivity pneumonitis) is a hypcrscnsitivity lung disease characterized by lymphocytic infiltrates in the pulmonary interstitial tissues. We have previously reported that the numbers of lymphocytes in bronchoalveolar lavage (BAL) samples in this disease correlate with levels of cholesterol and neutral lipid-laden “foamy” macrophages. We have also reported that the macrophages express an increased density of MHC class II antigens (in particular HLA-DQ) which are known to be essential for antigen recognition by T lymphocytes. The aim of the present study was to explore whether cholesterol is capable of enhancing the antigen-presenting function of mononuclear phagocytes by modulating the expression of HLA-D region products. Incubation of purified monocytes from healthy volunteers with cholesterol in scrum-free medium induced a significant increase in both the percentages of monocytes expressing HLA-DQ (P<0.02) and in the intensity of expression of the three HLA-D sub-region products. HLA-DQ. -DP and -DR (P<0.02, <0.01, <005. respectively). The cholesterol pre-incubated monocytes also exhibited enhanced antigen-presenting function (P<0.05), compared with controls pre-incubatcd without cholesterol. These findings indicate that increases in cholesterol in the extracellular milieu may augment antigen presentation by modulating the expression of HLA-D region products on antigen-presenting cells. Apart from EAA. this observation may also have relevance to inflammatory mechanisms in atherosclerosis, where‘foamy’ macrophages also occur in association with hypercholestcrolaemia.
Anti-P30 IgA antibodies as prenatal markers of congenital toxoplasma infection
Authors: DECOSTER A , DARCY F , CARON A , VINATIER D , DE L'AULNOIT D HOUZE , VITTU G , NIEL G , HEYER F , LECOLIER B , DELCROIX M , MONNIER J C , DUHAMEL M , CAPRON A
Published: July 2010

This study extends a previous study and confirms that the detection of anti-P30 IgA antibodies is very helpful in the diagnosis of acute acquired or congenital toxoplasmosis. Moreover, we demonstrate that an anti-P30 IgA response can be mounted in the fetuses infected by Toxoplasma gondii during their inira-utcrine life as early as week 23 of gestation. A double-sandwich ELISA described in our previous work was used to detect anti-P30 IgA antibodies in 1378 human serum samples collected from 551 patients, including 162 fetuses whose mothers had been infected by T. gondii during pregnancy, 46 congenitally infected and 90 uninfected newborns and 253 women suspected of having been infected during pregnancy, including the mothers of fetuses and newborns previously described. Anti-P30 IgA antibodies were detected in all cases of acute toxoplasmosis but in no case of chronic toxoplasmosis: in the majority of cases, the IgA antibody litre fell below cut-off in 3–9 months. Among the 46 congenitally infected newborns, anti-P30 IgA antibodies were detected in sera of 41 infected newborns (38 at birth, two in the first months of life, one in the seventh month of life), while anti-P30 IgM antibodies were detected in only 30 cases at birth and in one case during the first month of life. Among 162 fetuses, anti-P30 IgA response was observed in five infected fetuses, but was not detected in either 152 uninfected fetuses or in five fetuses considered as infected. The absence or presence of anti-P30 IgA antibodies in the fetus is discussed in relation to the date of maternal infection and collection of the fetal blood. It clearly appears from our study that the combined testing of both IgM and IgA in the fetus and the newborn is essential for a more efficient diagnosis of infection.
Involvement of HLA class II molecules in acquisition of staphylococcal enterotoxin A-binding activity and accessory cell activity in activation of human T cells by related toxins in vascular endothelial cells
Authors: UCHIYAMA T , ARAAKE M , YAN X J , MIYANAGA Y , IGARASHI H
Published: July 2010

Human umbilical vascular endothelial cells (HUVEC) express HLA class II molecules upon stimulation with recombinant human interferon-gamma (IFN-γ). Staphylococcal enterotoxin (SE) A (SEA)-binding assay using [125I]-SEA showed the presence of specific SEA binding in HUVEC stimulated with IFN-γ but not in unstimulated HUVEC. Levels of HLA class II expression and SEA-binding increased as the IFN-γ concentration and the period of stimulation were increased. Binding of [1254I]-SEA to the IFN-γ-stimulated HUVEC was reduced markedly by an anti-DR/DP MoAb. T cells produced IL-2 upon stimulation with a group of SEs (SEA, SEB, SEC, SED and SEE) in the presence HUVEC stimulated with IFN-γ but not in the presence of control HUVEC. The level of accessory cell activity in the IFN-γ-stimulated HUVEC was related to the level of HLA class II expression and SEA-binding activity. Antibodies to HLA class II molecules almost completely inhibited the response. These results indicate that HLA class II molecules are directly involved in the acquisition of these activities in HUVEC.
Induction and expression of protective T cells duringMycobacterium aviuminfections in mice
Authors: APPELBERG R , PEDROSA J
Published: July 2010

Mycobacterium avium is an opportunistic pathogen that infects individuals suffering from chronic lung disease or immunocompromised patients such as AIDS patients. Here we show that a highly virulent isolate of M. avium proliferated as extensively in T cell deficient as in immunocompetent mice. T cell deficient mice allowed a progressive growth of a less virulent AIDS-derived isolate of M. avium while immunocompetent mice arrested the growth of this isolate. Adoptive transfer of T cell enriched spleen cells between congenic strains of mice differing at the Bcg/Ity/Lsh locus showed that only naturally resistant BALB/c. Bcgr (C.D2) mice infected with the highly virulent strain of M. avium or the naturally susceptible BALB/c mice infected with the lower virulence isolate developed protective T cells and that these cells only mediated protection when transferred to naturally susceptible, but not to naturally resistant, mice. Both strains of M. avium proliferated in bone marrow-derived macrophages cultured in vitro and they were both susceptible to the bacteriostatic effects induced in the macrophages by crude lymphokines produced by concanavalin A-stimulated spleen cells.
Immune complex glomerulonephritis in experimental kala-azar II: Detection and characterization of parasite antigens and antibodies eluted from kidneys ofLeishmania donovani-infectedhamsters
Authors: SARTORI A , ROQUE-BARREIRA M C , COE J , CAMPOS-NETO A
Published: July 2010

In a previous report analysing kidney sections by immunofluorescence we showed that hamsters infected with L. donovani develop a glomerulonephritis (GN) associated with deposition of hamster immunoglobulins and parasite antigens in the glomeruli. In this study we characterize these immune components eluted from the kidneys. The eluted immunoglobulins showed specificity for L. donovani antigens and hamster immunoglobulins (rheumatoid factor-like activity). The four isotypes IgG1, IgG2, IgA and IgM were detected. Several L. donovani antigens were detected in the renal eluates by Western blot and immunoprecipitation using 125 I-labelled eluates. Proteins with mol. wt of 134, 82, 52, 31, and 26 kD were detected by Western blot and proteins with 134, 110, 93, 89 and 48 kD were detected by immunoprecipitation. With the exception of the 134 kD protein which was recognized by both rabbit anti-promastigote and rabbit anti-amastigote sera all the others were recognized only by the anti-amastigote serum. The 134 kD protein was the only one isolated from the kidneys of infected hamster immunocomplexed with IgG and was the only one detected in a promastigote lysate using IgG from L. donovani- infected hamsters.
Specific inhibition of mRNA accumulation for lymphokines in human T cell line Jurkat by mycobacterial lipoarabinomannan antigen
Authors: CHUJOR C S N , KUHN B , SCHWERER B , BERNHEIMER H , LEVIS W R , BEVEC D
Published: July 2010

The immunomodulatory effect of Mycobucterium tuberculosis-derived lipoarabinomannan (LAM) on mitogen/antigen-induced expression of mRNAs for a number of cytokines in human monocytic cell line Mono-Mac-6 and in T cell line Jurkat was investigated. Interestingly, LAM exhibited a down-regulatory effect on the accumulation of mRNAs for IL-2, IL-3, granulocyle-macrophage colony-stimulating factor (GM-CSF), and IL-2 receptor alpha (IL-2Rα) in T cells co-stimulated with phytohaemagglutinin-P (PHA) and 4β-phorbol-12-myristyl-13-acetate (PMA). In human Mono-Mac-6 cells, LAM has a weak inhibitory effect on the lipopolysaccharide (LPS)-induced mRNA accumulation for IL-1β, a slight stimulatory effect on mRNAs accumulation for IL-8 and tumour necrosis factor-alpha (TNF-α). but clearly no effect on mRNA accumulation for intercellular adhesion molecule-1 (ICAM-I). These findings imply that LAM may contribute to the immunologic defects associated with a number of mycobacterial infections by modulating these mediators.
Combined total deficiency of C7 and C4B with systemic lupus erythematosus (SLE)
Authors: SEGURADO O G , ARNAIZ-VILLENA A , IGLESIAS-CASARRUBIOS P , MARTINEZ-LASO J , VICARIO J L , FONTAN G , LOPEZ-TRASCASA M
Published: July 2010

The first inherited combined total deficiency of C7 and C4B complement components associated with SLE is described in a young female. Functional C7 assays showed a homozygous C7 deficiency in the propositus and her sister, and an heterozygous one in their parents. C4 molecular analyses showed that both the propositus and her mother had two HLA haplotypes carrying only C4A-spccific DNA sequences and a normal C4 gene number. Thus, only C4A proteins could be expressed, with resultant normal C4 serum levels. The coexistence of a combined complete C7 and C4B deficiency may therefore abrogate essential functions of the complement cascade presumably related to immune complex handling and solubilization despite an excess of circulating C4A. These findings challenge the putative pathophysiological roles of C4A and C4B and stress the need to perform both functional assays and C4 allotyping in patients with autoimmune pathology and low haemolytic activity without low serum levels of a classical pathway complement component.
Decreases in αβ T cell receptor negative T cells and CD8 cells, and an increase in CD4 +CD8+ cells in active Hashimoto's disease and subacute thyroiditis
Authors: IWATANI Y , AMINO N , HIDAKA Y , KANEDA T , ICHIHARA K , TAMAKI H , MATSUZUKA F , FUKATA S , KUMA K , MIYAI K
Published: July 2010

We examined peripheral lymphocyte subsets in patients with autoimmune thyroid disease, or subacute thyroiditis, in the active stage when possible. During destructive thyrotoxicosis arising from aggravation of Hashimoto's thyroiditis, both the numbers and proportions of αβ T cell receptor (TCR) negative T (WT31−CD3+) cells and CD8 (CD4−CD8+) cells decreased and those of CD4+CD8+ cells increased slightly, resulting in proportional increases in CD4 (CD4+CD8−) cells. non-T, non-B (CD5−CD19−) cells, and the CD4/CD8 cell ratio. Changes were similar in active subacute thyroiditis. During stimulative thyrotoxicosis in active Graves' disease, the numbers of such T lymphocyte subsets were not changed, but only the number of CD5+ B (CD5+CD19+) cells increased markedly, resulting in proportional decreases in total T (CD3+) cells, αβ+ TCR T (WT31+CD3+) cells, CD8 cells, and non-T, non-B cells. A serial study of some of the patients showed opposite changes in αβ TCR− T cells, the CD4/CD8 cell ratio, and CD5+ B cells between the active stages of Graves' and Hashimoto's diseases. αβ TCR− T cells were mostly γδ TCR+ T (IIF2+CD3+) cells in these patients. These data suggest that αβ TCR T (γδTCR+ T), CD8, and CD4+CD8+ cells are important in thyroid destruction in Hashimoto's disease and subacute thyroiditis, and that CD5+ B cells are important in thyroid stimulation in Graves' disease.
Analysis of different protein kinase C-dependent events in T cells from allogeneic bone marrow transplantation recipients
Authors: BALBOA M A , IZQUIERDO M , SÁNCHEZ-MADRID F , FERNÁNDEZ-RAÑADA J M , LÓPEZ-BOTET M
Published: July 2010

In order to understand the mechanisms underlying the T lymphocyte dysfunction associated to allogeneic bone marrow transplantation (BMT), we assessed two different protein kinase C (PKC) dependent events in T cells from BMT recipients: the PKC-dependent membrane expression and function of the CD69 early activation antigen; and the rapid phorbol ester-induced phosphorylation of PKC protein substrates in lysates from T cells permeabilized with digitonin, in the presence of (γ-32P)ATP. Most BMT recipient T cells detectably expressed the CD69 surface antigen after 24 h of stimulation with either phorbol 12-myristate 13-acetate (PMA) or anti-CD3 MoAb and PMA, thus indicating that PKC activity is sufficient to induce de novo gene expression. Nevertheless, it is noteworthy that the fluorescent staining intensity with anti-CD69 MoAbs was significantly lower in BMT recipient T cells than in normal T lymphocytes, although no clear-cut correlation was found between the expression of CD69 and the proliferative capacity. However, the pattern of PMA-induced phosphoproteins analysed as early as 1 min after PKC activation in T cells from BMT recipients displaying a low response to mitogenic stimuli, was undistinguishable from that detected in T cells from healthy subjects. In all cases a major 110-kD phosphoprotein was observed, which was inducible with PMA. phorbol 12,13-dibutyrate (PDBu). 1-oleoyl-2-acetylglycerol (OAG) and a phorbol-ester-related activator of PKC (mezerein); moreover, its phosphorylation was blocked by pretreating cells with the PKC inhibitor H-7. Altogether our results suggest that the depressed mitogenic responses, which were also observed in the present study when T cells were stimulated via CD69, cannot be simply attributed to a defective PKC activity.
Vasoactive intestinal peptide stimulates immunoglobulin production and growth of human B cells
Authors: ISHIOKA C , YOSHIDA A , KIMATA H , MIKAWA H
Published: July 2010

The effect of vasoactive intestinal peptide (VIP) on human lymphoblastoid B cell lines and tonsil B cells was studied. VIP increased immunoglobulin production and proliferation by lymphoblastoid B cell line. GM-1056, in a dose-dependent manner. As little as 10-12 M of VIP was effective, and higher concentrations of VIP induced an approximately five-fold increase in IgA production. Moreover, this enhancement was blocked by VIP antagonist. Similarly, VIP enhanced IgM and IgG production by other lymphoblastoid B cell lines, CBL and IM-9, respectively. In contrast to VIP, another neuropeptide substance P (SP) or somatostatin failed to enhance immunoglobulin production and thymidine uptake. VIP also enhanced IgA production and thymidine uptake by purified tonsil B cells. However, in contrast to B cell lines, VIP failed to enhance IgM and IgG production by tonsil B cells. SP or somatostatin failed to enhance immunoglobulin production or thymidine uptake by tonsil B cells. These results indicate that VIP acts as B cell stimulatory factor and that VIP may also have preferential effect on IgA production on tonsil B cells.
Characterization of neutrophil-mediated degradation of human C-reactive protein and identification of the protease
Authors: SHEPHARD E G , KELLY S L , ANDERSON R , FRIDKIN M
Published: July 2010

Human C-reactive protein (CRP) is shown to mediate a release of enzymatic activity from neutrophils which promotes its own degradation in the extracellular medium. This egress of proteolytic activity, which was upregulated by phorbol 12-myristate 13-acetate (PMA), was found to occur from both the cytoskeleton and membrane fractions of neutrophils and was dependent on the time of incubation of CRP with the cells and the concentration of CRP. Neutrophil kinases activated by PM A are found to be involved in upregulating the activity of the CRP-degrading protease. The apparent molecular weight of the CRP-degrading protease associated with the conditioned medium from PMA-stimulated neutrophils and neutrophil membrane and cytoskeleton preparations, was found by size exclusion chromatography to be 600 kD and migrated on 3–13% SDS-PAGE as four discrete bands to positions corresponding to apparent molecular weights of 209 kD, 316 kD, 398 kD and 501 kD.
Human trophoblast cells express CD4 and are permissive for productive infection with HIV-1
Authors: DAVID F J E , AUTRAN B , TRAN H C , MENU E , RAPHAEL M , DEBRE P , HSI B L , WEGMAN T G , BARRE-SINOUSSI F , CHAOUAT G
Published: July 2010

The European collaborative study of HIV-infected pregnant women in Europe now indicates a 13% risk of fetal HIV infection (originally thought to be about 30%, and possibly higher in some countries). Several reports suggest trans-placental passage. However, the detailed mechanisms associated with such vertical transmission have not yet been clarified. We have examined the possibility that HIV enters placental tissue from maternal blood via binding to CD4 and Fc receptors (FcR) at the trophoblast level, allowing intraplaccntal infection. Here we report the detection of several FcR with distinct localization in the placental villus as well as CD4 surface expression on human trophoblast cells. In addition, we show that trophoblastic cells interact specifically with the gp120/gp160 viral envelope protein. By their tissue localization, these receptors could be responsible for the entry of HIV into the fetal placental cells. Furthermore, purified placental cells can be directly infected by HIV in vitro, and the infection is inhibited by soluble CD4. This suggests a crucial role of the CD4 receptor but an additional way of entry cannot be excluded. Such an in vitro model may be suitable for further studies concerning placental HIV transmission and its prevention.
CD4 lymphocyte subset abnormalities associated with impaired delayed cutaneous hypersensitivity reactions in patients with X-linked agammaglobulinaemia
Authors: CROCKARD A D , BOYD N A M , McNEILL T A , McCLUSKEY D R
Published: July 2010

Circulating CD4 lymphocyte subset (CD45RA; CD45RO; CD29; Leu8) levels were determined in nine patients with -inked agammaglobulinaemia (XLA), nine patients with common variable immunodeficiency (CVI) and in 18 age– and se-atched controls. CD4CD45RO and CD4CD29 cells were significantly lower (P<001) in the XLA patient group (CD45RO, 15.7·10.2%; CD4CD29, 32.1 ±14.6%) compared with CVI patients (61.8·2504%; 6001 ±11.2%) and normal controls (43.7·22.3%, 54.5·22.0%). The levels of CD4CD45RA and CD4Leu8 cells were not abnormal in the XLA patient group. No selective reduction in CD4 subsets was observed in the CVI patient group. Delayed cutaneous hypersensitivity testing of five XLA and five CVI patients revealed a significantly reduced response to recall antigens in patients with XLA. This may relate to the deficiency of circulating memory T cells observed in these patients.
Differential expression of lymphocyte function-associated antigen (LFA-1) on peripheral blood leucocytes from individuals with Down's syndrome
Authors: BARRENA M J , ECHANIZ P , GARCIA-SERRANO C , ZUBILLAGA P , CUADRADO E
Published: July 2010

We analysed the expression of lymphocyte function-associated antigen LFA-1 on the cell surface peripheral blood lymphocytes, monocytes and granulocytes from 20 children wilh Down's syndrome. No differences in LFA-1 expression was found within monocytes or granulocytes from either normal or Down's syndrome children: however, a clear-cut difference was observed on lymphoid cells: Both normal and Down's syndrome lymphocytes displayed a bimodal pattern of LFA-1 staining by flow cytometry, with a predominance of cells with low expression in normal population, and an increased proportion of lymphocytes with high level of LFA-1 expression in Down's syndrome children. This difference correlates well with the abnormal proportion of T cell subsets and inversion of CD4/CD8 observed in a majority of our cases, and therefore, it could merely reflect the increase of certain T cell subsets nonnaily expressing higher number ofLFA-1 molecules. Taken together, our results do not support an abnormally increased expression of leucocytes integrins in trisomy 21 cells, and raise some doubt about the suggested role of the abnormal cellular expression of LFA-1 in the pathogensis of secondary immunodeficiency associated to Down's syndrome.
Liver-infiltrating T helper cells in autoimmune chronic active hepatitis stimulate the production of autoantibodies against the human asialoglycoprotein receptorin vitro
Authors: LÖHR H , TREICHEL U , PORALLA T , MANNS M , ZUM BÜSCHENFELDE K H MEYER , FLEISCHER B
Published: July 2010

Autoantibodies against the human asialoglycoprotein receptor (ASGPR) occur in the sera orpaticnts with autoimmune liver disorders. Live-nfiltrating T cell clones that specifically recognize the ASGPR have been described in patients with autoimmune chronic active hepatitis (A-AH) and primary biliary cirrhosis (PBC). Recently, we have shown that peripheral blood mononuclcar cells (PBMC) from patients with A-AH or PBC but not chronic viral hepatitis secreted ant-SGPR antibodies in vitro. In this study we characterized the influence of live-nfiltrating T cells on the secretion of ASGP-pecific autoantibodies by autologous B cells in cell culture supernatants. T cell clones from liver biopsies of three patients with chronic autoimmune liver disorders (one with A-AH, two with PBC) were isolated and investigated for their proliferative response to soluble ASGPR and their helper function provided to autoantibod-ecrcting B lymphocytes. PBMC from these patients secreted autoantibodies spontaneously in their cell culture supematants and showed a proliferative response to ASGPR. T cel-epleted PBMC, however, lacked spontaneous antibody secretion. Four CD4+ CD8− live-nfiltrating T cell clones showed a proliferative response to ASGPR and also induced spontaneous ant-SGPR antibody production in cell culture supernatants when added to autologous T cell depleted PBMC. Activated supernatants of these T cell clones failed to induce antibody production. None of seven CD4+CD8− and two CD4−CD8+ T cell clones no-esponding to ASGPR provided this help for antibody secretion. Ant-SGPR secretion in vitro could not be inhibited by the addition of MoAbs raised against monomorphic determinants on HLA class II molecules. The addition of purified ASGPR or polyclona-ctivating pokeweed mitogen showed no influence on the production of autoantibodies in these cultures. These data show that B lymphocytes require T cell help for the production of ASGP-pecific antibodies. This help can be provided by ASGP-esponsive T helper cells via cellular interactions.
Immunization of mice with the antigen A60 oi Mycobacterium bovis BCG
Authors: HUBBARD R D , FLORY C M , COLLINS F M , COCITO C
Published: July 2010

Antigen 60 (A60) is a thermostable component of the cytoplasm of Mycobacterium tuberculosis and BCG which can be fractionated into at least 15 protein bands when analysed by Western blot. Normal B6D2 mice were immunized subcutaneously with 20 μg of the A60 protein suspended in Freund's incomplete adjuvant (FIA) or in saline. Three weeks later the mice received a second dose of vaccine followed 2 weeks later by an aerogenic challenge with approximately 103 CFU of M. tuberculosis Erdman. The mice receiving the adjuvanled A60 showed a significant reduction (P < 0·05) in the number of viable organisms recovered from the lungs and the spleen 3 weeks after challenge. However, this response was less than that seen in BCG vaccinated controls.
B cell responses to HIV and the development of human monoclonal antibodies
Authors: BOYD J E , JAMES K
Published: July 2010

In this review B cell responses in HIV-infected individuals are summarized together with the techniques used to date to produce human monoclonals to HIV and the properties of these antibodies. Profound disturbances in B cell responses are apparent both in vivo and in vitro. While there is evidence in vivo of marked polyclonal B cell activation, primary and secondary antibody responses are impaired. Similarly these cells exhibit spontaneous immunoglobulin secretion upon in vitro culture but do not readily respond to B cell mitogens and recall antigens including HIV. Furthermore, certain of these defects can be reproduced in normal B cells in vitro by incubation with HIV or HIV coded peptides. Individuals infected with HIV develop antibodies to HIV structural proteins (e.g. p17, p24, gp41 and gp120) and regulatory proteins (e.g. vif, nef, RT). Autoantibodies against a number of immunologically important molecules are also frequently observed. The anti-HIV antibodies are predominantly of the IgG1 isotype and exhibit a variety of effects on the virus in vitro. To date, using conventional immortalization strategies, an appreciable number of human monoclonals to HIV have been developed. These have been specific for gp41, gp120 and gag with antibodies of the former specificity predominating. The majority of these antibodies have been of the IgGl isotype. Only a small number of the antibodies neutralize virus in vitro and most of these react with gp120. The neutralizing antibodies recognize conformational and carbohydrate epitopes or epitopes in amino acid positions 306–322. The predominant epitopes recognized by the anti-gp41 antibodies were in amino acid positions 579–620 and 644–662. A high percentage (≏ 25%) of these antibodies enhance viral growth in vitro. The problems relating to the production of human monoclonals to HIV are discussed together with strategies that could be used in the future.
Treatment of patients with chronic granulomatous disease with recombinant human interferon-gamma does not improve neutrophil oxidative metabolism, cytochrome b558 content or levels of four anti-microbial proteins
Authors: MÜHLEBACH T J , GABAY J , NATHAN C F , ERNY C , DOPFER G , SCHROTEN H , WAHN V , SEGER R A
Published: July 2010

Recombinant interferon-gamma (rIFN-γ) has been described to enhance phagocyte functions in vitro and in vivo in several patients with chronic granulomatous disease (CGD). To demonstrate the clinical usefulness of this treatment. 128 patients were treated in a randomized, double-blind multi-centre study with a placebo preparation or with rIFN-γ. We analysed parameters of neutrophil oxidative and non-oxidative metabolism in 16 patients enrolled in this study. No enhanced superoxide-release was observed in patients treated with rIFN-γ compared to placebo-treated patients. Phagocyte cytochrome b558 content also remained unchanged. Levels of four nonoxidative antimicrobial proteins (cathepsine G. azurocidine, p29b, lactoferrin) rose, fell, or remained unchanged, irrespective of treatment with rIFN-γ or placebo.
IL-6 in malignant pleural effusions and its augmentation by intrapleural instillation of IL-2
Authors: YANAGAWA H , SONE S , MUNEKATA M , ATAGI S , NII A , OGURA T
Published: July 2010

The levels and activities of endogenous IL-6 in malignant pleural effusions due to lung cancer before and during daily intrapleural instillations of recombinant IL-2 were examined by enzyme immunoassay and bioassay using an IL-6-dependent murine hybridoma cell line MH60.BSF2. Before therapy, malignant pleural effusions contained various levels of IL-6. Daily intrapleural instillation of recombinant IL-2 for treatment of malignant pleurisy resulted in significant augmentation of the levels and activities of IL-6 in the pleural effusions. On fractionation of the pleural effusion by chromatography, one major peak of material with a mol. wt of 24 kD showed IL-6 activity. In contrast, no significant level of tumour necrosis factor-alpha or IL-1β was detectable in pleural effusions before or during local IL-2 therapy. These data suggest that IL-2 is an important regulatory factor of secondary IL-6 production.
T and B cell reactivity to adrenal antigens in autoimmune Addison's disease
Authors: FREEMAN M , WEETMAN A P
Published: July 2010

Autoimmune Addison's disease is a rare condition of uncertain pathogcncsis. To delineate potential autoantigens. an adrenal homogenate was fractioned by SDS-gel clectrophoresis and used in immunoblotting and T cell proliferation assays. Antibodies specific for adrenal proteins with approximate molecular weights of 70, 55 and 45 were found in five out of 20 consecutive Addison's disease patients at routine follow-up. Five sera also reacted with a 52-kD protein shared with liver. T cells from six out of 10 Addison's disease patients proliferated in response to a range of adrenal-specific antigens including one, in particular, with a molecular weight of 18–24 kD. T and B cell reactivity to adrenal antigens did not appear to correlate and there was no relationship with time since diagnosis, associated autoimmunity or HLA-DR type. These results show that the autoimmune response to adrenal antigens in Addison's disease is heterogeneous and that such autoreactivity can only be inconsistently documented using these techniques and circulating antibodies or T cells.
Monocyte heterogeneity in angiotensin-converting enzyme induction mediated by autologous T lymphocytes
Authors: RYU J H , VUK-PAVLOVIC Z , ROHRBACH M S
Published: July 2010

The ability of monocyte subpopulations to be induced selectively by T lymphocytes to synthesize enhanced levels of angiotensin-converting enzyme (ACE) was examined using an in vitro model employing normal peripheral blood monocytes and T lymphocytes. Separation of monocytes into subpopulations on the basis of buoyant density indicated no difference in the ability of the resulting monocyte subpopulations to produce basal levels of ACE when cultured in the absence of T lymphocytes. However, the subpopulations differed significantly in their ability to synthesize enhanced levels of ACE in response to the presence of autologous T lymphocytes; low-density monocytes were induced by T lymphocytes to synthesize three-fold more ACE than were highdensity monocytes. Surface antigen labelling using MoAbs demonstrated that the low-density monocyte subpopulations also had a significantly higher percentage of Leu-M2+ monocytes compared with the high-density monocyte subpopulations. When monocytes were separated on the basis of the presence of the Leu-M2 antigen using an immune rosetting technique, T lymphocytes were able to induce significantly elevated levels of ACE in the Leu-M2+ enriched monocyte subpopulation but were unable to induce ACE beyond basal levels in the Leu-M2+-depleted monocyte subpopulation. These results demonstrate that monocytes are heterogeneous with respect to their ability to be induced by T lymphocytes to synthesize ACE. This raises the possibility that selective accumulation of a monocyte subpopulation in the granulomatous inflammation of sarcoidosis may be one of the factors required for elevated ACE synthesis in the resulting granuloma epithelioid cells.
Involvement of platelet-activating factor and tumour necrosis factor in the pathogenesis of joint inflammation in rabbits
Authors: ZARCO P , MAESTRE C , HERRERO-BEAUMONT G , GONZÁLEZ E , GARCIA-HOYO R , NAVARRO F J , BRAQUET P , EGIDO J
Published: July 2010

We have studied the participation of platelet-activating factor (PAF) in antigen-induced arthritis in rabbits, as well as the possible co-operation between PAF and tumour necrosis factor (TNF) in their ability to induce joint inflammation when injected into the knees of healthy rabbits. The administration of two structurally different PAF receptor antagonists, BN52021 and Alprazolam, from 4 h before the intra-articular injection of ovalbumin in preimmunized rabbits, induced an important reduction in the synovial fluid volume, in the amount of cells infiltrating the articular cavity and the synovial membrane, as well as in the prostaglandin E2 (PGE2) concentration. Furthermore, proteoglycans of the articular cartilage, which were found diminished in animals with non-treated arthritis, were well preserved in rabbits treated with PAF antagonists. All the synovial fluids from joints with arthritis had detectable amounts of PAF. The injection of either TNFor PAF into the joints of normal rabbits induced a mild inflammation. When TNF was administered 1 h before PAF, a synergistic response was noted in the synovial fluid volume, in the accumulation of leucocytes, and in the amount of PGE2. The administration of BN50726, a hetrazepine with a potent PAF-receptor antagonist effect, induced a diminution in those parameters. Our results suggest that PAF may be an early and important mediator of joint damage, and that TNF can amplify the inflammatory response induced by PAF. PAF receptor antagonists could play some role in the treatment of inflammatory joint diseases.
A human monoclonal autoantibody to a nucleolar structure
Authors: GONZALEZ M F , WICHMANN I , YELAMOS J , MELERO J , MAGARIÑO R , SANCHEZ-ROMAN J , NUÑEZ-ROLDAN A , SANCHEZ B
Published: July 2010

Peripheral blood lymphocytes from a scleroderma patient (CDC) were isolated, transformed with Epstein-Barr virus and fused to the heteromyeloma SHM-D33. Supematants from cultures were screened for autoantibody production against nucleoprotamine by ELISA. Positive wells were cloned by limiting dilution. After cloning, supematants from two wells were positive for the nucleoprotamine assay. One named CDC-1 has been studied in our laboratory. CDC-1 recognized a nucleolar antigen by indirect immunofluorescence. By using an ELISA with purified recombinant antigens. CDC-1 reacted against Ro/SS-A, Ul (RNP) and Sm. By immunoblotting using a lysate of MOLT-4 cell line, CDC-1 was able to react against a structure of 60 k D. When the antigen recognized by CDC-1 was purified, SDS-PAGE under reducing conditions with purified antigen and subsequent silver staining of the gel allowed us to delect three bands at 60,55 and 39 kD, respectively. A screening by ELISA with previously characterized antisera against our purified antigen demonstrated reactivity of the CDC-1 antigen with those antisera able to recognize Ro/SS-A.
Phospholipid-containing toxic malaria antigens induce hypoglycaemia
Authors: TAYLOR K , BATE C A W , CARR R E , BUTCHER G A , TAVERNE J , PLAYFAIR J H L
Published: July 2010

Hypoglycaemia is associated with severe malaria and is an important prognostic indicator. Molecules liberated during overnight incubation of erythrocytcs infected with Plasmodium yoelii induce marked hypoglycaemia in normal mice, with a delayed lime course compared with insulin: some, though weaker, activity could also be obtained by overnight incubation of uninfected crythrocytes. The active component shares many properties with (he phospholipid-containing molecules which we have previously shown to be toxic and to induce the release of tumour necrosis factor (TNF) from macrophages. However a MoAb which neutralizes the cytotoxicity of tumour necrosis factor in vitro did not prevent this induction of hypoglycaemia, whereas antiscrum against the toxic antigens did, as did immunization of normal (but not the immunoglobulin-deficient SCID) mice with the same material. Furthermore, normal mice injected with the antigens after immunization with phosphatidyl inositol or inositol monophosphale did not develop hypoglycaemia; the latter compound was also inhibitory when mixed with the antigens before injection. These compounds were previously shown to block the induction of TNF by the antigens and to induce the production of inhibilory antibodies. The role of these molecules in the etiology of the hypoglycaemia of malaria is discussed.
Anti-endothelial cell antibodies in nephropathia epidemica and other viral diseases
Authors: WANGEL A G , TEMONEN M , BRUMMER-KORVENKONTIO M , VAHERI A
Published: July 2010

Increased capillary permeability is a central feature of the severe forms of haemorrhagic fever with renal syndrome (HFRS) and occurs also, though less frequently, in ncphropaihia epidemica (NE). one of the milder forms of this syndrome, caused by Puumala virus. We therefore searched for anti-cndothclial cell antibodies (AECA) in patients with NE and in those with other presumed or serologically proven acute viral illnesses. By enzyme immunoassay. using human umbilical vein endothelial cells (HUVEC) as the substrate. IgG class AECA were detected significantly more frequently in patients with NE and with influenza A than in Red Cross blood donors. A lesser degree of reactivity could be shown with a human alveolar cell carcinoma line and with human and mouse embryonic fibroblasts. Prelreatment of HUVEC with interferon-gamma (IFN-y). but not with II.-I or tumour necrosis factor-alpha (TNF-a), increased their ability to bind IgG of sera from patients with NE and acute febrile illnesses. We conclude that, although AECA can be demonstrated in NE. they occur also in other acute viral illnesses and. unless cytopathic by a mechanism not shared by the AECA of these other illnesses, are unlikely to be causally related to the capillary leak in HFRS.
Plasma concentration of IL-6 in systemic lupus erythematosus; an indicator of disease activity?
Authors: SPRONK P E , ter BORG E J , LIMBURG P C , KALLENBERG C G M
Published: July 2010

To investigate the possible role of IL-6 in the activation of the autoimmune process in systemic lupus erythematosus (SLE), we serially measured concentrations of IL-6, IgG, and anti-dsDN A antibodies before and during exacerbations in patients with SLE. In addition, we serially related the IL-6 response to the generation of the acute phase reactant C-reaclive protein (CRP). Sixteen consecutive patients who developed an exacerbation were analysed in this study. Blood samples were drawn in EDTA monthly. At the time of maximal disease activity during exacerbation, IL-6 plasma concentrations were increased (≥6 pg/ml) in 12 out of the 16 cases. Concentrations of IL-6 correlated with the concentrations of CRP(P<0·01) and the score of the disease activity index (P <0·05). No correlation was found between IL-6 concentrations and concentrations of anti-dsDNA or IgG. The course of changes in IL-6 concentrations before the exacerbation was variable. Five out of the 16 exacerbations studied were characterized by a prominent rise of IL-6 at the time of maximum disease activity. In this subgroup serositis as well as elevated concentrations of CRP were observed more frequently (P<0–02). Seven exacerbations were not accompanied or preceded by changes in IL-6 concentrations and showed generally low IL-6 concentrations. In this latter subgroup cerebral involvement was seen more frequently (P<0·02). Our data do not suggest a pathogenic role for IL-6 in the generation of IgG and/or anti-dsDNA antibodies before exacerbations. Rises of IL-6 concentrations before exacerbations in SLE seem only to occur in a subgroup of patients with SLE characterized by the presence of serositis and elevated concentrations of CRP during exacerbation.
Control of immune-mediated disease of the central nervous system requires the use of a neuroactive agent: elucidation by the action of mitoxantrone
Authors: BAKER D , O'NEILL J K , DAVISON A N , TURK J L
Published: July 2010

Mitoxantrone was used as an immunosuppressive probe to elucidate a means for the control of experimental allergic enccphalomyelitis (EAE) induced in Biozzi AB/H mice following injection of spinal cord homogenate emulsified in Freund's adjuvant. A single i.p. injection of 2·5 mg/kg of mitoxantrone, 1–2 days before the anticipated onset of EAE, failed to prevent the majority of animals from developing clinical disease, whereas when the compound was injected directly into the central nervous system (CNS), at this time point, significantly increased therapeutic benefit was evident, with most animals failing to develop clinical EAE. Although the clinical use of intrathecal mitoxantrone is strongly contraindicated, these data suggest that increased therapeutic benefit may be achieved in immune-mediated disease of the CNS by targeting immunosuppressive doses of suitable agents, on lymphocyte activation within the CNS. In addition, direct administration of immunosuppressive doses into the CNS may reduce potentially unwanted (side) effects in the periphery.
Killing of human leukaemia/lymphoma B cells by activated cytotoxic T lymphocytes in the presence of a bispecific monoclonal antibody (αCD3/αCD19)
Authors: HAAGEN I A , van de GRIEND R , CLARK M , GEERARS A , BAST B , de GAST B
Published: July 2010

Bispecific antibodies (BsAb) can be used to retarget T cells irrespective of their specificity to certain target cells inducing target cell lysis. We have tested the efficacy of the BsAb SHR-1, directed against the T cell antigen CD3 and the B cell antigen CD 19 to induce (malignant) B cell kill by T cells as measured in a 51Cr-release assay. Two cytotoxic T cell clones (CTL). expressing TCRαβ or TCRγδ were effective in killing CD19 expressing B cell lines at different stages of differentiation in the presence, but not in the absence, of the BsAb. CDI9 target cells were not killed. Fresh CDI9* leukaemia/lymphoma cells were also efficiently killed by SHR-1 preincubated CTL clones. In addition, phytohaemagglutinin (PHA) or CD. Vactivatcd IL-2 expanded peripheral blood mono-nuclear cells (PBMC) of normal donors did so after 2 weeks of stimulation. A concentration of 100 ng/ml of the BsAb was sufficient to obtain optimal lysis of all target cells tested. These results show that fresh human leukaemia/lymphoma cells, freshly derived from active lymphoblastic leukaemia (ALL) as well as non-Hodgkin's lymphoma (NHL) patients, can be effectively killed in the presence of this BsAb by activated T cells.