bluuebunny/crossref_metadata_2025 huggingface dataset:
Complement, complement activation and anaphylatoxins in human ovarian follicular fluid
Authors: PERRICONE R , DE CAROLIS C , MORETTI C , SANTUARI E , DE SANCTIS G , FONTANA L
Published: July 2010

Functionally active complement was sought and detected in human follicular fluids obtained during the pre-ovulatory period. All the functional complement activities tested, including total haemolytic complement, classical pathway activity and alternative pathway activity were present in nine fluids from four different donors with values within the normal serum range. The immunochemical analysis demonstrated the presence of complement factors from C1 to C9, of B and of C1 INH, H, I. Complement anaphylatoxins were found employing RIA techniques in amounts significantly higher than in human plasma, thus demonstrating that follicular fluid complement, at least during the preovulatory period, is partially activated. A possible role for urokinase-like substances in such an activation was indicated by further in vitro experiments. The presence of active complement in follicular fluid can be relevant for the function of the enzymatic multi-factorial mechanism of ovulation.
Extracorporeal immunoadsorption therapy on rats. In vivo depletion of specific antibodies
Authors: NILSSON R , LINDGREN L , LILLIEHORN P
Published: July 2010

A technique for extracorporeal immunoadsorption of circulating specific antibodies from the plasma of rats is described. Catheterization of rats was performed using the carotid and the jugular blood vessels. The rats were treated non-anaesthetized. Blood was pumped continuously through a hollow-fibre plasma filter at a rate of 1.5 ml/min and plasma was separated and passed through an adsorbent column at a flow of 0.2 ml/min. The treated plasma was mixed with the blood cells before being returned to the rat. The column contained the antigen covalently linked to agarose beads. About three plasma volumes were treated during a period of 3 h and 30 min, which resulted in an antibody depletion of about 90–95%. The antibody levels returned to pre-adsorption levels after 4–5 days, or in some cases even exceeded the titres before treatment.
Antibodies to the peptide from the plasmid-coded Yersinia outer membrane protein (YOP1) in patients with ankylosing spondylitis
Authors: TSUCHIYA N , HUSBY G , WILLIAMS R C
Published: July 2010

Seventy-five Norwegian patients with ankylosing spondylitis (AS) were studied for class-specific antibody response against synthetic peptide, P81, representing the sequence of plasmid-coded outer membrane protein of Yersinia (YOP1) containing four amino acid homology (TDRK) with HLA-B27 sequence. Ten (16.7%), five (8.3%) and seven (11.2%) of 60 male AS patients showed elevated anti-YOPI P81 antibody of IgA, IgG, and IgM class, respectively, whereas for each isotype only one (4%) of 25 healthy male controls was positive. Differences were not observed between female patients and controls. In all isotypes, antibody-positive patients were more frequently found in patients with active disease. The anti-YOP1 P81 antibody levels of the patients were generally not correlated with the antibody levels against the peptide representing the hypervariable region of HLA-B27 (B27 peptide). However, in one patient the antibody was shown to react with both peptides by cross-inhibition analysis. Overall, it appears that any causal relationship between YOP1 and pathogenesis of AS is not strong. Immunogenicity and cross-reactivity of the YOP1 region encompassing the TDRE sequence particularly at the T cell level require further study.
HLA class II glycoprotein expression by monocytes from HIV-infected individuals: differences in response to interferon-gamma
Authors: ZWILLING B S , DARROW R , MCPEEK M , LAUFMAN H , PEARL D
Published: July 2010

The expression of HLA class II glycoproteins by monocytes from HIV-infected individuals evaluated. We found no differences in HLA-DR. HLA-DQ, or HLA-DP expression by freshly isolated monocytes from patients with clinical AIDS. HIV-infected patients without AIDS or from uninfected individuals. However, monocytes from HIV-infected individuals without AIDS responded better to stimulation with recombinant human interferon-gamma (IFN-γ) than did monocytes from patients with clinical AIDS or from uninfected individuals. The response to IFN-γ stimulation by monocytes from patients with clinical AIDS was different. Thus, the largest increases in the percentage of cells expressing HLA-DR was associated with the smallest changes in the fluorescence intensity of the population that is observed following stimulation of monocytes from other patient populations.
Rotavirus induces proliferative response and augments non-specific cytotoxic activity of lymphocytes in humans
Authors: YASUKAWA M , NAKAGOMI O , KOBAVASHI Y
Published: July 2010

In vitro cell-mediated immune responses to rotavirus in humans were studied. Peripheral blood mononuclear cells (PBMC) of healthy adults proliferated in response to stimulation with the infectious and u.v.-inactivated Wa strain of human rotavirus, showing a maximum response on day 7 of culture; however, cord blood lymphocytes failed to respond to rotavirus. A Cross-reactive proliferative response of PBMC detected by stimulation with the NCDV strain of bovine rotavirus suggests the existence of epitopes common to both human and bovine rotaviruses. which are recognized by human T lymphocytes. The phenotype of the majority of activated lymphocytes was CD3+4+8-, indicating that the cells mainly activated were helper T cells. Culture supernatants of PBMC stimulated with rotavirus contained interleukin-2 (IL-2) and interferon-gamma (IFN-γ). In addition, PBMC stimulated with rotavirus demonstrated significantly enhanced cytotoxic activity against natural killer (NK) sensitive K562 ceils as well as an NK-resistant Epstein-Barr virus-immortalized lymphoblastoid cell line (LCL). Treatment of PBMC with anti-CDl6 or NKHIA monoclonal antibody, both of which react with most NK cells and lymphokine-activated killer cells and complement markedly reduced the cytotoxic activity against K562 and LCL. These results suggest that stimulation of human PBMC with rotavirus results in the production of tymphokines, such as IL-2 and IFN-γ, by rotavirus-reactive helper T cells and that these lymphokines augment NK activity and generate other forms of non-specific cytotoxic human lymphocyte activity. These cell-mediated immune responses observed in the present in vitro study might play an important role in protection and recovery from rotavirus infection.
Lymphokine-activated killer cytotoxicity against leukaemic blast cells
Authors: TRATKIEWICZ J A , SZER J , BOYD R L
Published: July 2010

While the sensitivity of cell lines and solid tumours to lymphokine-activated killer (LAK) cell cytotoxicity is extensively reported, the ability of these generated cytotoxic ccMs to lyse leukaemic blast cells is controversial. This study reports the successful generation of LAK cells with specificity for leukaemic blast cells. Control donors, patients in remission and patients with active leukaemia were capable of lysing allogeneic leukaemic blast cells. One patient was also capable of lysing autologous leukaemie blast cells. Successful LAK generation was achieved by the use of high dose recombinant interleukin-2 (rIL-2) while combinations of lymphokines did not improve LAK specificity or efficacy. These findings suggest that LAK immunotherapy may be considered for the treatment of patients with acute leukaemia.
Monoclonal antibodies to early pregnancy factor perturb tumour cell growth
Authors: QUINN K A , ATHANASAS-PLATSIS S , WONG T-Y , ROLFE B E , CAVANAGH A C , MORTON H
Published: July 2010

The pregnancy-associated substance early pregnancy factor (EPF) has previously been reported as a product of tumours of germ cell origin. More recently EPF (or an EPF-related substance, tEPF) has also been detected in the serum of patients bearing tumours of non-germ cell origin. We report here the production of tEPF by a variety of cultured transformed and tumour cell lines, of both germ and non-germ cell origin. Antibodies specific for EPF remove all tEPF activity from tumour cell conditioned medium, tEPF production is found to be associated with cell division; tEPF is no longer detected after growth arrest or differentiation. Co-culture of tumour cells with increasing doses of anti-EPF monoclonal antibodies resulted in a significant, dose-dependent decrease in rate of cell growth and viability. Similar anti-EPF concentrations had no effect on the concanavalin A induced proliferation of mouse spleen cells. These studies suggest, therefore, that tEPF is a growth-regulated product of cultured tumour and transformed cells. These cells are also dependent upon tEPF for continued growth, i.e. tEPF is acting in the autocrine mode.
Characterization of normal human CD3+ CD5- and γδ T cell receptor positive T lymphocytes
Authors: SROUR E F , LEEMHUIS T , JENSKI L , REDMOND R , FILLAK D , JANSEN J
Published: July 2010

The functional and phenotypic properties of normal human CD3+ CD5- T cells which have a higher frequency of cytotoxic cells than CD3+CD5+ T lymphocytes have been described. Using three- and four-colour immunofluorescence flow cytometric cell sorting, the CD3+CD5- and CD3+CD5+ populations were subdivided into αβ or γδ T cell receptor positive cells. The four subsets were examined for the in vitro cytotoxic activity and were also stimulated with mitogens in limiting-dilution assays to measure the frequencies of proliferating and interleukin-2 (IL-2) producing cells. CD3+CD5-αβ, CD3+CD5-γδ+ and CD3+CD5+γδ+ cells had lower frequencies of proliferating and IL-2-producing cells than did CD3+CD5+αβ+ cells. However, the cytotoxic activity of the different phenotypes was higher in the CD3+CD5- subsets, especially when these cells were γδ+ Expression of γδ or lack of expression of CD5 appeared to be associated with the acquisition of cytolytic potentials. CD8 was expressed on 20% of fresh CD3+γδ+ cells. Cultured γδ cells retained the expression of γδ, but quickly lost that of CD8 and with time modulated the expression orCD5, The expression of CD5 was found to be higher on sorted CD3+CD5+γδ- than on CD3+CD5+γδ+ cells. These observations indicate that γδ is preferentially expressed on CD5-negative or weakly positive T lymphocytes and that CD3+CD5-γδ cells appear to constitute a discrete small subset of mature T lymphocytes which are cytotoxic in nature. However, the exact immunological function of these cells and their place in T cell ontogeny are yet to be elucidated.
Seasonal increase of spontaneous histamine release in washed leucocytes from rhinitis patients sensitive to grass pollen
Authors: WEYER A , GUINNEPAIN M T , SUTRA J P , BORGNON A , HERPIN-RICHARD N , ICKOVIC M R , MEAUME J , RAFFARD M , TEKAIA F
Published: July 2010

The spontaneous histamine release (SHR) in basophils from patients sensitive to grass pollen has been studied before and during the 1987 grass pollen season. Nineteen patients were recruited on seasonal rhinitis symptoms, positivity for cutaneous tests and for serum-specific IgE with grass pollen. At the time of the biological investigations the patients were following a clinical trial of hyposensitization, including placebo, calcium phosphate and aluminium hydroxide-adsorbed grass pollen extract treatments. During the pollen season, grass pollen counts and clinical scores were checked over a 40-day period. Mean SHR was significantly higher during the pollen period than before, for the whole population of 19 patients (10.9% and 4.6%; P< 0.005) as well as when the high SHR responders were-excluded (5.5% and 3.6%; P< 0.01). No significant correlation existed between SHR and clinical scores or treatments. SHR could be inhibited at 4°C, in absence of Ca++ or of oxidative metabolism and thus originated from cells actively secreting histamine.
S protein binds to serum-treated agarose beads independently of complement activation and the formation of the terminal complement complex on the beads
Authors: HETLAND G , GARRED P , PETTERSEN H B , MOLLNES T E , JOHNSON E
Published: July 2010

Comparison of initial fearly-phased and terminal (late-phase) sequence activation of complement by agarose beads and endotoxin was evaluated in an enzyme immunoassay (EIA) of serum levels of C3c and C9 neoepitopes. Respectively. EIA and Western boloting with anti-S protein monoclonal antibody revealed lower S protein values and weaker S protein bands in serum activated by agarosebeads than by endotoxin implaying that S protein was removed from serum by binding to agarose. The binding of S protein to the beads was confirmed by radioimmunoassay and was found to be equal m normal and heat-inactivated serum. In contrast, the terminal complement complex was formed only on agarose heads invubated with normal serum and not with inactivated serum.
Molecular mimickry between HLA B27 andYersinia, Salmonella, ShigellaandKlebsiellawithin the same region of HLA α1-helix
Authors: LAHESMAA R , SKURNIK M , VAARA M , LEIRISALO-REPO M , NISSILÄ M , GRANFORS K , TOIVANEN P
Published: July 2010

Two new examples of amino acid homology between HLA B27 and microbes triggering HLA B27-associated diseases are described. An outer membrane protein YadA (Yersinia adhesin, previously called Yop1) of Yersinia enterocolitica and Y. pseudotuberculosis shares a linear tetrapeptide with HLA B27. A cationic outer membrane protein OmpH of Salmonella typhimurium shares homology with five amino acids of HLA B27 in a non-linear fashion. The four amino acids of YadA are also notably included in the hexapeptide identical between Klebsiella pneumoniae nitrogenase and HLA B27, and three of them occur in the pentapeptide shared by a Shigella flexneri protein and HLA B27. Antibodies against synthetic peptides including HLA B27 homologous sequences of YadA and OmpH were observed in one-third of the patients with HLA B27 associated diseases. Antibodies were directed against a flanking sequence next to the amino acid sequences shared by arthritis-triggering microbes and HLA B27. The area of identity in each example of this molecular mimicry (Yersinia, Salmonella, Shigella and Klebsiella) is located in the same place on the H LA B27 molecule: between amino acids 70 to 78 in the variable region of α1-helix. This area of HLA B27 molecule includes sites predicted to be important for binding processed antigens.
Regulation of experimental autoimmune uveitis in rats—separation of MHC and non-MHC gene effects
Authors: HIROSE S , OGASAWARA K , NATORI T , SASAMOTO Y , OHNO S , MATSUDA H , ONOÉ K
Published: July 2010

Experimental autoimmune uveitis (EAU) is an organ-specific autoimmune disease and has served as a model of certain ocular inflammatory conditions in man. The present study was aimed at separating the effects of MHC and non-MHC genes on the development of EAU in the rat. EAU-susceptible LEW (RT1l) EAU-resistant WKAH (RTIk), and WKAH.1L (RTF) MHC congenic strain of WKAH background rats were immunized with retinal soluble antigen (S-Ag) in Freund's complete adjuvant (FCA). LEW rats showed typical EAU, while neither WKAH nor WKAH. 1L congenic rats developed EAU. However, when an additional i.v. injection of Bordetella pertussis was given, all rat strains developed EAU. Furthermore, when immunized with peptide M, an 18-mer synthetic peptide, which corresponds to amino acid positions 303–320 of bovine S-Ag, and given an additional i.v. injection of B. pertussis, LEW and WKAH. IL rats developed EAU, whereas WKAH did not. When ACI (RTIavl), BUF (RTIb), LEJ (RTIl), W(RTIk), F344(RTI1v1), BN (RTIq), NIG-III (RTIq), TO (RT1I), and SDJ (RT1u) rats were immunized with peptide M or S-Ag and then given B. pertussis, all strains developed EAU by immunization with S-Ag plus B. pertussis, but only F344 and NIG-III developed EAU by immunization with peptide M. These findings suggest that susceptibility to EAU in rats is controlled by both MHC and non-MHC genes; and that in the absence of B. pertussis adjuvant, the form of disease induced by native S-Ag in FCA is governed by non-MHC gene(s). However, this effect of non-MHC gene(s) could no longer be observed when the rats were also injected with B. pertussis adjuvant at sensitization.
Variant CD45R expression with autosomal dominant inheritance affects both helper/inducer (CD4 +) and suppressor/cytotoxic (CD8 +) T cell populations
Authors: SCOTT C S , RICHARDS S J , COLE J C , GOOI H C
Published: July 2010

The differential expression of various membrane CD45R isoforms by normal lymphocyte populations is known to be closely associated with distinct immunoregulutory functions. Abnormal patterns of CD45R expression have been reported in patients with common variable immunodeficiency and HIV infection, and recent evidence has suggested the possibility that one type of variant CD45R expression may be inheritable. By multiple colour flow cytometry, we studied the immunological characteristics of CD4+ helper/inducer and CD8+ suppressor/cytotoxic T cells in a family wilh variant CD45R expression over three generations. This variant pattern of CD45R expression was shown to affect both CD4+ and CD8+ lymphocyte populations in individual family members and was immunologically characterized by a failure of the normal reciprocal expression of the CD45RA and CD45RO isoforms. Family studies also revealed that this trait had an autosomal dominant mode of inheritance and. in the heterozygous state, appeared not to be associated with major clinical abnormalities. The different isoforms of CD45 show distinct patterns of expression during lymphocyte ontogeny and activation, and these patterns appear to closely reflect function. Despite their diverse immunological roles, our finding that both CD4+ and CD8+ T cells in affected family members showed the same defect suggests a common regulatory mechanism(s) for both these lymphocyte populations. The recognition of this abnormality, particularly in homozygous individuals, will be of great importance in understanding the role of these molecules in immune function and disease.
Anti-idiotype and immunosuppressant treatment of murine lupus
Authors: MORLAND C , MICHAEL J , ADU D , KIZAKI T , HOWIE A J , MORGAN A , STAINES N A
Published: July 2010

The effect of the administration of a xenogeneic anti-idiotype antibody (anti-Id33) to a cross-reactive idiotype (Id33) present on anti-dsDNA antibody was examined in 6-week-old (NZB/NZW) F1 (BWF1) female mice. The administration of anti-Id33 led to a transient reduction in immunoglobulins expressing Id33, followed by a rise at 30 and 34 weeks that was significantly higher than in untreated mice (P<0.05). Likewise, anti-dsDNA antibody levels were significantly higher at 10 and 18 weeks than in untreated mice (P<0.0l). No differences were seen in survival to 40 weeks, proteinuria or the severity of glomerulonephritis. Concurrent administration of cyclosporin A(CyA) with anti-Id33 markedly ameliorated glomerular injury and proteinuria and improved survival. By contrast, glomerular injury, proteinuria and survival were worse in mice treated with cyclophosphamide plus anti-Id33, compared with untreated mice. Neither CyA nor cyclophosphamide treatment, when given with anti-Id33 altered serum levels of anti-dsDNA, anti-ssDNA or Id33+ immunoglobin, compared with untreated mice. The different effects of CyA and cyclophosphamide on T lymphocytes and their discrepant effects on glomerular injury when given with anti-Id33 in this model lead us to postulate a role for T lymphocytes in the glomerular injury of BWF1 lupus.
Hyper IgE in stimulatory graft-versus-hostdisease: role of interleukin-4
Authors: DOUTRELEPONT J M , MOSER M , LEO O , ABRAMOWICZ D , VANDERHAEGEN M L , URBAIN J , GOLDMAN M
Published: July 2010

Intravenous injection of 2 × 108 DBA/2 spleen cells into adult intact (C57BL/6 x DBA/2) Fl mice results in a stimulatory graft-versus-host reaction (GVHR) linked to the recognition by donor CD4+ T cells of Ia alloantigens on host B cells. In the experiments presented here, we found that this GVHR is associated with a major increase in IgE serum levels which was already present 7 days after the cell transfer. At 6 weeks, mean IgE levels were more than 200-fold above the control values. Host B cells were responsible for the hypersecretion of IgE in stimulatory GVHR since it was also observed when the DBA/2 donor inoculum was depleted of B cells but not when the Fl recipients were irradiated. The induction of IgE secretion required donor CD4+ T cells as treatment of the donor inoculum with lytic anti-CD4 monoclonal antibody (MoAb) completely prevented the occurrence of the hyper IgE whereas depletion of CD8+ cells had no influence on this parameter. The role played by interleukin-4 (IL-4) in this model was analysed in vivo by the administration of the 11B11 anti-IL-4 rat MoAb (total dose 36 mg) during the first 12 days following induction of stimulatory GVHR by 8 × 107 DBA/2 spleen cells. This treatment completely prevented the development of hyper IgE whereas the administration of a control rat MoAb had no significant effect. We conclude that stimulatory GVHR in mice is associated with a major increase in serum IgE which is mediated by IL-4.
Lactoferrin-inducible monocyte cytotoxicity for K562 cells and decay of natural killer lymphocyte cytotoxicity
Authors: MCCORMICK J A , MARKEY G M , MORRIS T C M
Published: July 2010

Monocyte-enrichcd and lymphocyte-enriched fractions of peripheral blood from three healthy volunteers were obtained by percoll density gradient centrifugation. The cytotoxie activity of each fraction against 51Cr-labellcd K562 cells was quantified in a 2-h assay using freshly isolated cells of each fraction and cells of each fraction which had been incubated with and without lactoferrin in complete medium for 18 h before performing the assay. We have thereby shown that cytotoxicity was not demonstrable in the lymphocyte fraction (containing 7.3 ± 2% large granular lymphocytes) after 18 h in medium, whereas the cytotoxicity of the monocyte fraction (containing 3.0 ± 4% large granular lymphocytes) was still significantly increased (P≤ 0.01) and that lactoferrin had no effect on lymphocyte fraction cytotoxicity while producing an 11-fold increase in the cytotoxicity of the monocyte fraction. It is therefore possible to perform a relatively simple test of monocyte cytotoxicity using lactoferrin as a stimulant in a 2-h 51Cr-labelled K562 assay system by allowing 18 h to elapse for lymphocyte natural killer cytotoxicity to decay.
Characterization of immune inducer and suppressor macrophages from the normal human lung
Authors: SPITERI M A , POULTER L W
Published: July 2010

Monoclonal antibodies (MoAbs) that are able 10 discriminate between dendritic cells (MoAb RFD1+) and mature macrophages (MoAb RFD7+ (in normal tissues were used in combination with density separation techniques to isolate relatively homogeneous subpopulations of macrophages from human bronchoalveolar lavage (BAL). A characterization of surface antigen expression, and functional capacity was then carried out on each isolated alveolar macrophage (AM) subset. One population with the phenotype RFD1+ RFD7- obtained from the non-adherent cell pool showed the characteristics of antigen-presenting cells having absent or poor expression of Fc and C3b receptors, a low content of lysozomal hydrolase and poor phagocytic capacity. This population strongly stimulated T lymphocytes in allogeneic mixed lymphocyte reactions (MLR). A second AM population, isolated by adherence and density centrifugal ion expressed the phenotype RFD1+ RFD7+. These cells showed the same phenotypic characteristics of mature macrophages with strong expression of C3b and Fc receptors, and marked phagocytic capacity. Such AM were very poor stimulators of allogeneie MLR. Under certain circumstances the RFD1+ RFD7+ cells were shown to actively repress the stimulatory capacity of the RFD1+ RFD7- subpopulation. These results suggest that variations within the functional capacity of AM subsets may be capable of influencing the strength of acquired T cell immune responses of the lung.
Analysis of the spontaneousin vitroanti-HIV-1 antibody secretion by peripheral blood mononuclear cells in HIV-1 infection
Authors: VENDRELL J P , SEGONDY M , DUCOS J , REYNES J , HUGUET M F , NICOLAS J C , SERRE A
Published: July 2010

We studied the spontaneous in vitro secretion of anti-HIV-1 antibodies by peripheral blood mononuclear cells (PBMC) from HIV-1-infected patients. Specific antibody production was detected in supernatants of PBMC cultures using an ELISA; HIV-1 specificity was confirmed by antigen adsorption and Western blotting. This antibody secretion was found to be an active phenomenon and was not due to a release of plasma antibodies passively adsorbed onto the cell membranes. In all positive supernatants, anti-HIV-1-secreted antibodies were directed against enu-encoded antigens and many supernatants also contained antibodies to pol- and gag-encoded antigens. PBMC from all HIV-1-infected patients tested (140 adults and 18 infants) secreted anti-HIV-1 antibodies. This production was found during all the clinical stages of HIV-1 infection. Our results suggest that this spontaneous HIV-1-specific antibody secretion represents a marker of HIV-1 infection. Detection of these antibodies could be a valuable tool for early confirmation of HIV-1 infection in neonates born to HIV-1-seropositive mothers.
Host and bacterial factors control theMycobacterium avium-induced chronic peritoneal granulocytosis in mice
Authors: APPELBERG R , PEDROSA J M , SILVA M T
Published: July 2010

Persistent peritoneal granulocytosis and elevated macrophage counts have been found in nine mouse strains from 8 to 90 days after infection with Mycobacterium avium. Peritoneal granulocylosis was higher in M. avium-resistant BALB/c. Bcgr (C.D2) mice, compared with congenic M. avium-susceptible BALB/c (Bcgb) animals. Although maximal granulocytosis values were not related to virulence of the inocula, the kinetics of the granulocytic response varied with the virulence of M. arium. Following infections by avirulent (rough) strains of M. avium, the peritoneal granulocytosis progressively declined in BALB/c and C3H/He mice. A similar decline in granulocyte number was observed in resistant C3H/He mice infected with virulent M. avium (smooth transparent strain). In both instances the decline in the peritoneal granulocytosis was associated with a progressive elimination of the inoculum. In the susceptible BALB/c mice, virulent M. avium strains induced progressive infection accompanied with a rapid decline in granulocyte number, whereas the infection with attenuated M. avium, which caused a chronic infection, induced persistent granulocytosis. The ability to recruit granulocytes following the intraperitoneal inoculation of a phlogistic substance (casein hydrolysate) was decreased in infected susceptible but not in infected resistant mice at 90 days of infection with virulent M. avium.
Anti-nuclear antibodies of primary biliary cirrhosis recognize 78–92-kD and 96–100-kD proteins of nuclear bodies
Authors: FUSCONI M , CASSANI F , GOVONI M , CASELLI A , FARABEGOLI F , LENZI M , BALLARDINI G , ZAULI D , BIANCHI F B
Published: July 2010

The specificities of anti-nuclear antibodies (ANA) reacting with multiple nuclear dots (MND-ANA) present in about 15% primary biliary cirrhosis sera were studied by Western blot analysis with nuclear fractions from a human cell line. Reactivity with two broad bands of 78–92 kD and 96–00 kD of the insoluble fraction was present exclusively in MND-ANA-positive sera. Antibodies eluted from these proteins specifically retained the immunofluorescence reactivity of MND-ANA. Immunomorphological analysis by a pre-embedding technique revealed that the antibody specifically binds to nuclear regions resembling in size and number nuclear bodies. Since these structures are absent in immature rate endometrial cell and can be induced by diethylstilbestrol. we tested MND-ANA by immunofluorescence on cryostat sections of uteri from hormone-treated and untreated immature rats. A strong reaction of nuclear dots was observed predominantly in endometrial cells of hormone treated rats. We thus conclude that MND-ANA present in primary biliary cirrhosis sera are directed against 78–92-kD and 96–100-kD nuclear proteins located in nuclear bodies.
Expression of intercellular adhesion molecule-1 (ICAM-1) on thyroid epithelial cells in Hashimoto's thyroiditis but not in Graves' disease or papillary thyroid cancer
Authors: BAGNASCO M , CARETTO A , OLIVE D , PEDINI B , CANONICA G W , BETTERLE C
Published: July 2010

In order to study the possible role of antigen-independent adhesion systems in thyroid autoimmunity. we evaluated by indirect immunofluorescence the expression of lymphocyte functional antigen-1 (LFA-1) and its ligand ICAM-1 on mononuclear cell infiltrates (when present) and thyroid follicular cells of four patients with Hashimoto's thyroiditis, 30 with Graves' disease, five with papillary cancer, two with follicular adenoma, and two normal thyroid specimens. The expression of MHC class I and class II antigens was also evaluated. Most mononuclear infiltrates were LFA-1 positive, as expected. A positivity for ICAM-1 on follicular cells was observed in three out of four Hashimoto's thyroiditis specimens; such a phenomenon was totally absent in Graves' disease or any other pathological condition, or in normal tissue. MHC class II expression on thyrocytes was observed in all the patients with Hashimoto's thyroiditis, in 27 out of 30 with Graves' disease and in three out of five papillary cancer specimens.
Low CD4/CD8 T lymphocyte ratio in acute myocardial infarction
Authors: SYRJÄLÄ H , SURCEL H-M , ILONEN J
Published: July 2010

T lymphocyte subsets were analysed using monoclonal antibodies and flow cytometry to determine whether myocardial infarction and cardiopulmonary resuscitation induce changes in these. Groups of 11 infarct patients and 10 patients with past cardiopulmonary resuscitation were compared with 11 age- and sex-matched controls and 12 sepsis patients. The differences in the CD4/CD8 ratios between the four groups were significant (F= 7.71, P=0.001). The infaret patients had lower CD4/CD8 ratios (mean±s.d. 0.83.0.43) than the control (2.12.1.13; P=0.001) or sepsis cases (1.76.1.05; P=0.004), but their ratios did not differ from those of the resuscitation group (0.93±0.79, P=0.84). The latter group also had lower ratios than the control (P=0.003) and sepsis groups (P=0.013). Most infarct patients had an on admission inverted CD4/CD8 ratio which usually returned to normal in the next 2 days. A permanently low CD4/CD8 ratio may be a poor sign prognostically after both myocardial infarction and resuscitation.
Selective activation of T cells in newly diagnosed insulin-dependent diabetic patients: evidence for heterogeneity of T cell receptor usage
Authors: KONTIAINEN S , TOOMATH R , LOWDER J , FELDMANN M
Published: July 2010

Cell surface phenotyping of 58 newly diagnosed diabetic children and 25 controls confirmed the presence of activated T cells, expressing HLA class II antigens or receptors for interleukin-2 (1L-2R, CD25) in the majority or the patients. Some of these cells putatively include those involved in islet cell destruction, as reported previously. Monoclonal antibodies recognizing three families of the variable regions of the β chain (Vβ) of the T cell receptor were used to determine the percentage of peripheral blood cells expressing those specific gene segment products. The number of the activated T cells from each Vβ family was compared with that of the resting T cells of the same family in the patients and the controls. In 18 out of 58 (31%) of these patients there was evidence of oligoclonal proliferation of activated T cells as judged by marked increases in cells expressing a V/β family in the IL-2R+ T cell pool, compared with the total T cell pool. However, different V/β families were augmented in individual patients, indicating considerable heterogeneity of T cell activation in different patients. These results are in contrast to murine models of autoimmunity, where virtually monoclonal T cell activation, restricted to a single V/β family has been reported.
Common variable immunodeficiency is associated with polymorphic markers in the human major histocompatibility complex
Authors: HOWE H S , SO A K L , FARRANT J , WEBSTER A D B
Published: July 2010

Common variable immunodeficiency (CVI) is a heterogeneous condition characterized by arrest in B cell differentiation. A high frequency of null alleles of the C4 gene has been reported in patients with this disorder. We investigated the restriction fragment length polymorphisms (RFLP) of the MHC class II genes HLA-DRB, DQA, and DQB, the class III gene C4 and the tumour necrosis factoralpha (TNF-α) gene in 40 Caucasian patients. The results showed an increase in HLA-DR3 in patients (40%vs 30.5%), but, more significantly, there was a striking increase in the number of CVI patients who carried a deletion of the C4A gene (46%vs 25%). In both patients and controls there was strong allelic association between HLA-DR3 and C4A deletion, and HLA-DR3 and TNF-α. Our results suggest that genes present on an extended haplotype containing these three polymorphisms contribute to genetic susceptibility to CVI.
Blood lymphocyte proliferation, cytokine secretion and appearance of T cells with activation surface markers in cultures with Helicobacter pylori. Comparison of the responses of subjects with and without antibodies to H. pylori
Authors: KARTTUNEN R
Published: July 2010

A whole inactivated H. pylori bacterium preparation was found to stimulate blood mononuclear cells from both antibody-positive and antibody-negative subjects, but the antibody-positive subjects tended to have lower proliferation responses. The present study was designed to characterize T cell activation further by measuring several components of the response. Eighty-seven subjects (80 dyspeptic patients and seven healthy persons from the laboratory staff) with or without antibodies to H. pylori were studied by measuring the DNA synthesis induced by several H. pylori concentrations 1–23 μg/ml) and the control stimulants PPD, tetanus toxoid and pokcweed mitogen (PWM). H. pylori-induced secretion of interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-α), interleukin-4 (IL-4), soluble CD8 and IL-2 receptor (IL-2R) molecules and H. pylori- and PPD-induced appearances of IL-2R+ and HLA-DR+ T cells were measured in a smaller number of subjects. H. pylori-induced DNA synthesis was again lower in the antibody/bacterium-positive subjects, while no differences between the two groups were found in cultures stimulated by unrelated antigens or PWM. Soluble IL-2R and TNF-α were detectable in cultures with H. pylori from all subjects, while the amount of IL-2 did not differ from that in the background culture. No differences were found in the amounts of IL-2 or soluble IL-2R between the antibody-positive and negative subjects; while the former tended to secrete more soluble CD8 molecules, a difference which was significant with the smaller H. pylori concentration used (P<0.01). The numbers of HLA-DR+ and IL-2R+ T cells increased in cultures with H. pylori or PPD from all the subjects, the majority of both cells having the CD4 phenotype. Numbers of DR+ and IL-2R+ T cells were similar in the cultures of the antibody-positive and negative subjects, but the respective CD8 subsets were increased in the former. The confirmed decrease in proliferation in the antibody-positive subjects does not seem to be connected with lower IL-2/IL-2R responses but may involve CD8 cell activation.
Functionally active complement proteins C6 and C7 detected in C6- and C7-deficient individuals
Authors: WÜRZNER R , ORREN A , POTTER P , MORGAN B P , PONARD D , SPÄTH P , BRAI M , SCHULZE M , HAPPE L , GÖTZE O
Published: July 2010

Two sensitive sandwich ELISAs based on monoclonal antibodies directed to native C6 and C7 allowed the detection and quantitation of these complement proteins in 20 out of 37 serum samples from individuals who had previously been classified as deficient in these proteins as assessed by immunochemical and/or functional assays. Furthermore, serum from four C6-deficient and one combined C6-/C7-deficient individual showed an increase in the terminal complement complex (TCC) and a decrease in native C6 and C7 after complement activation as assayed by specific ELISAs. Despite their (incomplete) deficiencies, these individuals therefore possess functionally active terminal complement proteins with respect to their ability to generate the TCC. As these individuals have no history of a susceptibility to neisserial infections, even low concentrations of functionally active C6 and C7 may provide sufficient protection against those micro-organisms whose destruction requires TCC formation.
Characterization and large production of human monoclonal antibodies against the HIV-1 envelope
Authors: BOYER V , BROLY H , SOUCHE S , MADAULE P , ROSSIER J , ZAGURY D , DESGRANGES C
Published: July 2010

Peripheral blood lymphocytes from a volunteer immunized with a recombinant vaccinia virus VSC-25 expressing the gp160 env protein of HTLV-IIIB strain and from an asymptomatic HIV-infected individual were immortalized by Epstein-Barr (EBV). Clones which secrete human monoclonal antibodies from the two individuals (DZ, IgGl, λ and C31, IgG1, κ) were obtained and were stable for more than 2 years. The two monoclonals were directed against the gp 160 env protein of HIV, DZ directed against the gp41 and C31 directed against the gp120. C31 was group-specific, whereas DZ was directed against the HTLV-IIIB and HTLV-RF strains. The epitope recognized by DZ was mapped to the carboxy terminus of the gp41, by expression of HIV DNA fragments in a yeast system and peptide analysis. The C31 epitope was not expressed by the yeast library and not present among the peptides which were tested. Monoclonal antibodies had no inhibitory effect in an HIV-induced cell fusion assay, but DZ showed a weak neutralizing activity against the HTLV-IIIB strain. Cloned EBV-transformed cell lines were fused to a murine myeloma, which allowed the heteromyeloma to be cultivated in serum-free medium. The monoclonal antibodies were produced in large quantity in a hollow-fibre reactor at defined culture conditions and purification procedures.
Cytokine induction of neopterin production
Authors: HENDERSON D C , SHELDON J , RICHES P , HOBBS J R
Published: July 2010

The pteridine neopterin is a marker of immunological activation and has been shown to be a useful marker of graft-versus-host disease (GVHD) in bone marrow transplant patients. High levels of both neopterin and interferon-gamma (IFN-γ) were produced in vitro during mixed lymphocyte responses, which may be considered to be a model of the primary events leading to GVHD. Neopterin was shown to be produced by monocytes in response to stimulation with IFN-γ, but not other cytokines. However, the interleukins IL-lα, IL-1β, IL-2, and tumour necrosis factor (TNF) alpha and beta, but not IL-6, stimulated neopterin production by unfractionated peripheral blood mononuclear cells (PBMC), and culture supernatants from PBMC stimulated with IL-1 α, 1L-1 β, IL-2 and IL-6, but not TNF-α or TNF-β induced neopterin production following transfer to fresh monocyte cultures. It therefore appears that cytokines may generate neopterin by induction of IFN-γ, by synergy with low levels of induced IFN-γ, or by non-IFN-γ-dependent mechanisms.
Acute-phase protein synthesis in human hepatoma cells: differential regulation of serum amyloid A (SAA) and haptoglobin by interleukin-1 and interleukin-6
Authors: RAYNES J G , EAGLING S , MCADAM K P W J
Published: July 2010

Interlcukin-6 (IL-6, BSF-2 or IFN-β2) is thought to be the major regulator of the acute-phase protein response that follows tissue injury and inflammation, with interleukin-1 (IL-1), tumour necrosis factor and more recently, LIF or HSF III, slightly stimulatory on only certain acute phase proteins. The synthesis of the major acute-phase protein SAA, originally described as being synthesized in response to IL-1, has been claimed recently to be mainly under IL-6 regulation. Our results show that in the human hepatoma cell line HuH-7, IL-1 is the major stimulating cytokine increasing SAA synthesis by a factor in excess of 100-fold. We also show that under most conditions interleukin-6 and tumour necrosis factor stimulate additively in combination with IL-1. Isoelectric focusing has demonstrated that SAA1 and SAA2α are expressed but not SAA2β. The HuH-7 cell line is IL-6 responsive since haptoglobin is stimulated mainly by IL-6.
Interleukin-4 regulates the interleukin-2 receptors on human peripheral blood B lymphocytes
Authors: TOMIZAWA K , ISHIZAKA A , KOJIMA K , NAKANISHI M , SAKIYAMA Y , MATSUMOTO S
Published: July 2010

The high-affinity interleukin-2 (IL-2) receptor (IL-2R) consists of the non-covalent association of at least two subunits, p55 and p70–75, capable of binding IL-2 with low and intermediate affinity, respectively. We studied the effects of cytokines on the IL-2R expressed on human peripheral blood B lymphocytes using monoclonal antibodies specific for IL-2R p55 and IL-2R p70–75, by means of two-colour flow cytometric analysis. In freshly isolated peripheral blood B lymphocytes, the p55 subunit was expressed only in a small population (7.0% of CD20+ cells), whereas the p70–75 subunit was expressed in a large population (89.0% of CD20+ cells). Of the cytokines studied, interleukin-4 (IL-4) and interferon-gamma (IFN-γ) were involved in the regulation of IL-2R on B cells. After a 2-day incubation with IL-4, expression of IL-2R p55 was markedly induced, but expression of IL2-R p70 –75 was profoundly suppressed in a dose-dependent manner. These abilities of IL-4 to promote IL-2R p55 expression and suppress IL-2R p70–75 expression were inhibited by the presence of I FN-γ. Other cytokines, including IL-1, IL-2, IL-5, and IL-6, had little effect on the expression of these two subunits. These findings suggest that IL-4 is a cytokine modulating B cell response through the regulation of IL-2R.
Monocytes secrete factors regulating glycosaminoglycan synthesis in mesangial cellsin vitro
Authors: NARITA I , MORIOKA T , ARAKAWA M , SHIMIZU F , OITE T
Published: July 2010

The present study was conducted to determine the manner in which monocytes increase mesangial matrices, particularly glycosaminoglycans (GAGs) which interact with various other matrix components such as collagens, laminin, fibronectin and lipoproteins. A supernatant of human peripheral blood monocyte cultures activated by lipopolysaccharide (LPS) contains stimulating factors for glycosaminoglycan synthesis in rat mesangial cells (MCs). The culture supernatant in this study was concentrated and fractionated by gel chromatography and the GAG-stimulatory factor was found to have a molecular weight of 10–17 kD. This factor was shown to be present in fractions different from that of IL-1. Gel and ion-exchange chromatography studies of GAGs synthesized by MCs indicated the elution patterns of GAGs in the presence and absence of the monocyle culture supernatant to be essentially the same. Local infiltration of monocytes into the glomerulus. often seen in various types of glomerular injury, may be an important factor in the accumulation of the mesangial matrix.
Quantification of cross-reactive idiotype-positive rheumatoid factor produced in autoimmune rheumatic diseases. An indicator of clonality and B cell proliferative mechanisms
Authors: SHOKRI F , MAGEED R A K , KITAS G D , KATSIKIS P , MOUTSOPOULOS H M , JEFFERIS R
Published: July 2010

The aetiology of sustained autoantibody production in human autoimmune diseases is unknown. Evidence for structural similarities and common clonal origin among autoantibodies have been demonstrated through the expression or cross-reactive idioiype (CRI). In the present study we use four monoclonal antibodies (MoAbs) with specificity for non-overlapping CRI on human rheumatoid factor (RF) autoantibodies to define the structural features of polyclonal RF characteristic of patients with autoimmune rheumatic diseases. The pattern of CRI expression in the scrum of 12 patients with rheumatoid arthritis (RA), eight with systemic lupus crythcmalosus(SLE) and 20 with primary Sjögren’s syndrome and 34 normal individuals were determined in parallel with the level of igM RF. IgA RF and autoantibodies to the cellular antigens SS-A. SS-B. Sm. nRNP and dsDN A and cryoglobulins. The results demonstrate significant elevation in the level of IgM and IgA expressing VHI (G6 and G8) and YHl 11 (B6 and Dl2) associated CRI in the scrum of patients with autoimmune rheumatic diseases compared with normal individuals. These increases paralleled, but did not equal the increase in the level of immunoglobulins and RF. However, when expressed as proportion of immunoglobulin. only the VHI-associated CRI were significantly elevated in patients compared with normal individuals. The proportion of IgM RF expressing the VMI -associated CRI was higher in patients with Sjdgrcn’s syndrome compared with SLE and RA. Furthermore, the proportion of IgA RF expressing the G6 CRI was higher than G66 IgM RF. These findings imply that different mechanisms contribute to RF production in autoimmune diseases. It is suggested thai polyconal B cell activation is likely to be a contributing mechanism. However, such polyclonal activation is unlikely to be random since a selective elevation in the level of specific autoantibodies and VMI -associated CRI is observed. Furthermore, the data demonstrate that a proportion of autoanlibodies in autoimmune diseases arc immunoglobulin gcrmline gene encoded. This is more evident in some patients with primary Sjogrcn’s syndrome, where RF is likely to be oligoclonal or monoclonal in individuals with lymphoprolifcration.
Contribution of the spleen, lymph nodes and bone marrow to the antibody response in collagen-induced arthritis in the rat
Authors: RAHMAN J , STAINES N A
Published: July 2010

The relative contribution of different lymphoid tissues to the anti-CII antibody response was studied in rats with arthritis produced as a result of immunizing them with collagen type II (CII). Antibody production was measured by maintaining lymphoid cells in short-term culture in collagen-coated microculture wells: the antibody they secreted was determined directly by a modified ELISA. Systemic sensitization to CII was established within a week of immunization, and a stronger response in the local draining lymph nodes relative to the spleen was associated with the development of clinical disease. From experiments involving splenectomy and adoptive cell transfer, the spleen was ascribed a suppressive role in controlling both arthritis and total antibody production. The bone marrow was found to be an important site of antibody production and the greater production of antibody by cells from tibial marrow in limbs with arthritis, compared with healthy limbs, argues for a local immune response to degrading joint antigens that may have systemic suppressive or protective properties. It is concluded that local immunity reflects the state of disease and that the antibodies produced by different lymphoid tissues may be made in response to different stimuli, and that the antibodies in turn may have different pathological effects.