bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunogenicity of HIV-1LAI gp160 and env peptides in squirrel monkey Saimiri sciureus using alumine and experimental adjuvants
Authors: Perraut R , Chouteau P , Moog C , Bonnemains B , Kieny M P
Published: November 2003

Since the identification of the HIV virus, important advances have been achieved in the definition of potential subunit vaccines. We investigated the immunogenicity of a recombinant gp160 antigen and of two gp41 peptides from HIV-1LAI associated with seven different adjuvant formulations in squirrel monkeys. All animals were immunized twice with gp160 and then with a gp41 peptide using the same formulation. All adjuvants used led to a subsequent antibody response against gp160, and 55% of the animals immunized developed anti-gp160 antibodies that could neutralize the virus in vitro. Specific anti-gp41 antibody response was also observed. Results obtained underlined the key role of the adjuvant formulation in the antibody response against a given part of the immunogen, and indicate that such immunogenicity-related investigation can be carried out conveniently in the squirrel monkey Saimiri sciureus.
RNA-labelled Ro and La ribonucleoprotein complexes reassembled in vitro; characterization by gel shift analysis
Authors: Granger D , Gendron M , Tremblay A , Chabot B , Ménard H A , Boire G
Published: November 2003

Ro and La RNP complexes were reassembled from in vitro labelled hY5 RNA and HeLa cell extracts. These complexes were then visualized through retardation of migration of labelled hY5 RNA in non-denaturing polyacrylamide gels. Three major complexes (named A, B, and C) were formed when crude cellular extracts (S100 fraction) were used. Using monospecific anti-60-kD Ro (Ro60) and anti-La antibodies to retard RNPs containing these antigens during migration in the gels, the three major complexes were shown to contain Ro60 (C), La (B), or both proteins (A). The specificity of RNA–protein interactions in the reassembled complexes was further demonstrated using two 3′-shortened hY5 RNA transcripts lacking the La-binding site (hY5–Alu I RNA) and both the Ro60 and La-binding sites (hY5–Hha I RNA). hY5–Hha I RNA still formed a single, minor complex when incubated with S100 extract, suggesting interaction with a yet undefined protein. In addition, we used the capacity of specific antibodies to retard the migration of the ressembled complexes to design a detection assay for anti-Ro and anti-La autoantibodies. Using 84 human sera, our assay was shown to approximate the specificity and sensitivity of an immunoprecipitation assay where 32P-labelled cell extracts are used as source of antigens. Our assay may be used to detect low levels of antibodies to conformational determinants on Ro60 and La proteins in human sera and antibody preparations.
Antibody reactivity profiles following immunization with diverse peptides of the PERB11 (MIC) family
Authors: Leelayuwat C , Hollingsworth P , Pummer S , Lertmemongkolchai G , Thom G , Mullberg J , Witt C , Kaufman J , Degli-Esposti M A , Cosman D , Dawkins R
Published: November 2003

PERB11 (MIC) is a gene family possessing multiple copies located within the MHC. Structurally, PERB11 is related to the MHC class I, neonatal IgG Fc receptor (FcRn) and Zn-α2-glycoprotein molecules. The MHC class I family is complex in terms of its genomic arrangement, expression and function, and available evidence suggests that the PERB11 family may be similarly complex. We have adopted an approach to study the expression of such complex gene families by immunizing with multiple peptides and by screening the resulting antibodies against a large range of tissues. The amino acid sequences of PERB11.1 and PERB11.2 as well as those of other related molecules were analysed and compared. Peptides were chosen for immunization based upon (i) loop formation within the equivalent known structure of the MHC class I molecules; (ii) immunogenicity by computer analysis; and (iii) evolutionary relationships. Antibodies in serum from immunized rabbits bound to three out of six peptides used for immunization. ELISA and immunoprecipitation demonstrated binding both to the peptides and to the PERB11.2 recombinant protein. By immunofluorescent staining of various tissues of several species, the three antisera generated overlapping profiles of activity. These included reactions with kidney, small and large intestine, oesophagus, testis, ovary and human neutrophils. This is the first description of antibodies induced by the PERB11 peptides. The extreme complexity of these profiles requires further investigation, but may be explained in terms of antibodies against diverse products of the PERB11 gene family and/or related molecules.
Autoantibodies to double-stranded (ds)DNA immunoprecipitate 18S ribosomal RNA by virtue of their interaction with ribosomal protein S1 and suppress in vitro protein synthesis
Authors: Tsuzaka K , Winkler T H , Kalden J R , Reichlin M
Published: November 2003

We report that four systemic lupus erythematosus (SLE) patient sera containing anti-dsDNA antibodies, three affinity-purified anti-dsDNA IgG, and a human anti-dsDNA MoAb (33.H11) immunoprecipitate 18S ribosomal RNA from DNase-treated 32P-labelled MOLT4 cell extract. This 18S RNA precipitation was inhibited completely by preincubating 33.H11 with calf thymus dsDNA or the recombinant human ribosomal protein S1, which was reported to cross-react with anti-dsDNA antibodies (J Immunol 1996; 156:1668–75). Whole IgG from three SLE sera with anti-dsDNA antibodies, 33.H11, and three affinity-purified anti-dsDNA IgG inhibited in vitro translation of globin mRNA (percent inhibition was 36–50%). This translation inhibition by anti-dsDNA antibodies was enhanced (67–79%) when the reticulocyte lysate was treated with DNase. Suppression of protein synthesis could be a pathogenic mechanism of anti-dsDNA antibodies, since it has also been shown that anti-dsDNA penetrates living cells (J Immunol 1995; 154:4857–64) in culture.
Respiratory syncytial virus-induced cytokine production by neonatal macrophages
Authors: Tsutsumi H , Matsuda K , Sone S , Takeuchi R , Chiba S
Published: November 2003

The induction of immunoregulatory cytokines IL-1β, IL-6, IL-12, tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) was studied with neonatal (cord blood) monocyte-derived macrophages (MDM) after in vitro infection with respiratory syncytial virus (RSV). The expression of mRNAs for these cytokines in RSV-infected MDM was examined by reverse transcriptase-polymerase chain reaction (RT-PCR). The activities of these cytokines were assayed by ELISA. Significant increase of expression of mRNA for IL-6, IL-12, TNF-α and IFN-γ occurred within 2 h after infection and decreased within 6 h after infection. At 20 h after infection the MDM produced and secreted moderate levels of IL-6 and TNF-α; however, no IL-12 and IFN-γ activities were detected. Moderate IL-1β mRNA was expressed before RSV infection, and its expression increased at 2 h after infection. However, no detectable IL-1β was secreted in culture fluids. These observations suggest that RSV-infected neonatal macrophages produce and secrete IL-6 and TNF-α quickly during the eclipse phase of RSV infection and therefore may play a prominent role in the initiation of the immune response to RSV.
Expression of leucocyte–endothelial adhesion molecules is limited to intercellular adhesion molecule-1 (ICAM-1) in the lung of pneumoconiotic patients: role of tumour necrosis factor-alpha (TNF-α)
Authors: Vanhée D , Molet S , Gosset P , Tillie-Leblond I , Boitelle A , Wallaert B , Tonnel A-B
Published: November 2003

Coal workers' pneumoconiosis (CWP) is characterized by a chronic inflammatory lung reaction associated with macrophage accumulation in alveolar spaces. In this study, we investigated in CWP the implication of adhesion molecules such as E-selectin, ICAM-1 and vascular cell adhesion molecule-1 (VCAM-1) and the role of TNF-α which is one of the cytokines inducing their expression. Adhesion molecule expression was analysed by immunohistochemistry on lung biopsies from patients with CWP and from healthy subjects. In parallel, soluble adhesion molecules were detected in bronchoalveolar lavage fluids (BALF) from patients by specific ELISA. The involvement of TNF in the induction of these adhesion molecules was measured (i) by immunohistochemistry on sections from lung fragments, and (ii) by evaluating in vitro the expression of adhesion molecules on endothelial cells and on alveolar epithelial cells in the presence of alveolar macrophage supernatants. In control subjects, a weak staining of ICAM-1 was detected only in alveolar walls, while E-selectin and VCAM-1 were undetectable. In pneumoconiotic patients, ICAM-1 was expressed at a high level by endothelium, by alveolar and bronchial epithelial cells and by alveolar macrophages. E-selectin and VCAM-1 expression remained undetectable. Measurement of soluble adhesion molecule showed that only the concentration of sICAM-1 was significantly increased in BALF from patients with CWP compared with controls. The involvement of TNF in this ICAM-1 expression was shown by the in vitro effect of alveolar macrophage supernatants on adhesion molecule expresssion by endothelial cells and epithelial cells (this effect was neutralized by anti-TNF antibodies) and by the increased production of TNF in the lung of pneumoconiotic patients. These data provide evidence for the involvement of ICAM-1, induced at least in part by alveolar macrophage-derived TNF, in the development of the inflammatory reaction in CWP.
Characteristics of asymptomatic secondary immune responses to measles virus in late convalescent donors
Authors: HUISS S , DAMIEN B , SCHNEIDER F , MULLER C P
Published: November 2003

Among 44 fully protected, late convalescent adults re-exposed to measles, four developed an asymptomatic secondary immune response (SIR) with a significant increase in measles virus (MV)-specific IgG and low IgM. The boosted antibodies were mainly of the IgG1 subclass and reacted with the nucleoprotein and the haemagglutinin protein. About 30 weeks after re-exposure, antibody levels had decreased by 35–50%, suggesting that the booster effect may only be transient. SIR was only found in individuals with a pre-exposure IgG level below a well defined threshold. Antibody levels above this threshold fully protected against SIR. SIR seems to be an ‘all or none response’ where the magnitude of increase in specific IgG is independent of pre-exposure antibody levels as long as these are below the above threshold. In combination with pre-exposure neutralizing and haemagglutination inhibiting titres, a threshold was defined below which SIR is likely to occur. This may be useful to predict susceptibility to SIR in a given population, since individuals undergoing clinically inapparent SIR are among seropositive subjects, the most likely candidates to support transmission of virus.
Surface antigen expression in spleen cells of C57Bl/6 mice during ageing: influence of sex and parity
Authors: BARRAT F , LESOURD B M , LOUISE A , BOULOUIS H-J , VINCENT-NAULLEAU S , THIBAULT D , SANAA M , NEWAY T , PILET C H
Published: November 2003

So far all studies on the murine ageing process have been conducted on virgin mice. Immune ageing may be influenced by sex hormone differences related to sex or pregnancies. The aim of this study was to investigate whether pregnancies and gender influence the cell changes observed during ageing in a peripheral lymphoid compartment of C57Bl/6 mice. Using flow cytometry, changes in (Thy1.2+) T cell, (B220+) B cell and (CD11b/Mac-1) macrophage spleen populations were monitored in 2, 8 (3 months after last pregnancy) 15 and 23-month-old mice including males, virgin and multiparous females. The development of naive (CDCD44low), memory (CD44high), activated/memory (MEL-14, CD62L) cells were investigated in CD4+ and CE8+ T cell subsets. Both short term (at 8 months) and long term (at 15 and 23 months) effects of multiparity were obvious in the lymphocyte/macrophage population changes associated with the ageing process. Short-term effects included delayed appearance of CD4+CD44high memory lymphocytes and increased numbers of both CD4+MEL-14low activated/memory cells and Mac-1+ macrophages when compared with virgin control mice. Later effects of multiparity were increased CD8αdull populations and increased T/B cell ratios and the ratio of memory to naive CD4+ cells (CD44+high/CD44+low). A sex effect was noticed: males exhibited lower Mac-1+ levels and memory/naive ratio in CD4+ subset than virgin females throughout life. These results suggest that gender and/or pregnancies affect the age-related distribution of lymphoid and macrophage cell populations in the spleen of C57Bl/6 mice.
Characteristics of synovial fluid effusion in collagen-induced arthritis (CIA) in the DA rat; a comparison of histology and antibody reactivities in an experimental chronic arthritis model and rheumatoid arthritis (RA)
Authors: ERLANDSSON HARRIS H , LILJESTRÖM M , KLARESKOG L
Published: November 2003

We have characterized the cellular content and some antibody reactivities in synovial fluid (SF) from DA rats with CIA. Since CIA is widely used as a model for RA, in which many studies concerning immune responses are performed on SF samples, we considered it important to describe the local, disease-causing immune reactions in CIA. At the peak of disease (day 22 after immunization), the major cell population in CIA SF was granulocytes (72%), but macrophages (17·9%), plasma cells (2·6%) and lymphocytes (7·7%) were also present. The CIA synovial membrane (SM) obtained at the same time was mainly infiltrated by monocytes, with granulocytes, lymphocytes and plasma cells also present. Cell populations in blood did not differ between arthritic and normal DA rats. Equally, high anti-collagen type II (CII) and rheumatoid factor (RF) levels could be detected both in SF and in sera. Notably, RF levels were also increased in normal DA rats. Moderate levels of anti-heat shock protein 65 kD (hsp) antibodies were recorded systemically in both normal and diseased animals. In conclusion, the cellular composition in SF and in SM are similar in rat CIA and in RA. The morphological differences between SF and SM that are characteristic for RA could also be demonstrated in CIA. The antibody data indicate systemic production of anti-CII and anti-hsp antibodies as well as RF, but they give no support for local production of these antibodies in the joints, which is the case in RA.
Measurement of deoxyribonuclease I (DNase) in the serum and urine of systemic lupus erythematosus (SLE)-prone NZB/NZW mice by a new radial enzyme diffusion assay
Authors: MACANOVIC M , LACHMANN P J
Published: November 2003

A new radial enzyme diffusion (RED) method for the measurement of DNase activity in serum and urine is described. The sensitivity of the assay is in the range of 15.6–500 ng/ml. The assay is based on the hydrolysis of double-stranded (ds) DNA (or nucleosomes) in agarose. The specificity of the reaction for DNase I was established by showing that either EDTA in the reaction buffer or G-actin abolished DNase activity. Being a functional assay, RED has advantages over radioimmunoassay (RIA) or ELISA, since antigenic assays may also measure complexes of DNase with actin. This method was used to measure DNase activity in the sera and urine of lupus-prone mice (NZB/NZW F1 hybrids, aged 4–6 weeks). Serum DNase activity in these mice was significantly lower (mean 9 ng/ml) than in control, normal mice of the same age and sex (mean 37 ng/ml). Concentration of DNase in the urine of 4–6-week-old female NZB/NZW F1 hybrids (24 ng/ml) was significantly lower then in control mice (521 ng/ml). The RED method was used to measure the concentration of actin as the DNase inhibitor in serum. G-actin in the presence of ATP binds DNase and inhibits its nucleolytic activity. Since ATP is necessary for the actin inhibition of DNase I, this shows that there is actin as well as DNase I in the serum. Actin is not only ATP-dependent, but also heat-labile. Heating the sera for 10 min at 50°C increases DNase activity. This is an alternative method for measuring the concentration of actin in the serum. An almost identical estimate of actin concentration in sera of normal mice was found from the difference of DNase activity in the presence or absence of ATP (mean actin concentration = 21 ng/ml) or from the difference of DNase activity in heated and non-heated serum (mean actin concentration 18 ng/ml). We were not able to demonstrate DNase inhibitors in the urine of either control or NZB/W F1 hybrid mice.
Anti-ribosomal and ‘P-peptide’-specific autoantibodies bind to T lymphocytes
Authors: STAFFORD H A , CHEN A E , ANDERSON C J , PAUL A G A , WYATT E L , LEE L A , NEAS B R
Published: November 2003

Patients with systemic lupus erythematosus (SLE) frequently have anti-lymphocyte autoantibodies, some of which also bind to surfaces of neurons. Since anti-ribosomal P protein autoantibodies (anti-P) from SLE patients also bind to surfaces of neurons, we hypothesized that anti-P are anti-lymphocyte antibodies. A panel of human T lymphocytes was evaluated for anti-P binding by indirect immunofluorescence. Affinity-purified anti-ribosomal antibodies were used as a source of anti-P. These autoantibodies bound to the surfaces of all transformed T cell lines tested. This binding was not mediated by Fc receptors. It was inhibitable by ribosomes. Anti-P bound to circulating T lymphocytes from healthy adults and children. They also bound to thymocytes and cord blood T cells from normal neonates. Circulating T cells from SLE patients with anti-P bound less anti-P than cells from healthy controls. Two patients were studied on multiple occasions. The capacity of their T cells to bind anti-P correlated inversely with titres of anti-ribosomal antibodies. Anti-ribosomal antibodies, other than anti-P, also appear to bind to T cells. The surface of T cells contains a protein with the size and antigenicity of the ribosomal P protein, P0. We conclude that anti-ribosomal antibodies are a subset of anti-lymphocyte autoantibodies. Their possible role in the pathogenesis of lymphopenia or lymphocyte dysfunction in SLE has to be defined in further studies.
Heterozygous C8β complement deficiency does not predispose to meningococcal disease
Authors: PLATONOV A E , SHIPULIN G A , SHIPULINA O JU , VERSHININA I V , DENSEN P
Published: November 2003

We have identified 42 Russian patients with homozygous C8β complement component deficiency, all of whom had experienced at least one episode of systemic meningococcal disease. About 90% of these individuals have a C → T exchange in exon 9, leading to a premature stop codon. If, like the homozygous-deficient state, heterozygous C8β deficiency constitutes a risk factor for meningococcal disease, it would be expected to be detected with increased frequency among individuals suffering from this disease. Using allele-specific polymerase chain reaction (PCR)we studied 153 consecutive patients with meningococcal disease admitted to the Moscow Hospital for Infectious Diseases to determine the frequency of C8 null allele. No individuals with heterozygous C → T exchange were identified among these 153 patients, despite the fact that seven persons were detected who had homozygous C8β deficiency, caused by the same C → T exchange in exon 9, and one patient who had C7 component deficiency. Thus, heterozygous deficiency, although more frequent than homozygous deficiency in the general population, does not result in a substantial increase in susceptibility to meningococcal disease.
Hyperexpression of transporter in antigen processing-1 (TAP-1) in thyroid glands affected by autoimmunity: a contributory factor to the breach of tolerance to thyroid antigens?
Authors: SOSPEDRA M , TOLOSA E , ARMENGOL P , ASHHAB Y , URLINGER S , LUCAS-MARTIN A , FOZ-SALA M , JARAQUEMADA D , PUJOL-BORRELL R
Published: November 2003

According to the ‘aberrant HLA expression’ hypothesis, endocrine autoimmunity is driven by presentation of self antigens by target cells over-expressing HLA molecules. In autoimmune thyroid diseases (AITD), thyroid follicular cells (thyrocytes) over-express HLA class I and HLA class II molecules. Since efficient presentation of endogenous peptides via class I requires transporters that translocate endogenous peptides from the cytoplasm to the endoplasmic reticulum, i.e. transporters associated with antigen processing (TAP) -1 and -2, the capability of thyrocytes to express TAP and whether TAP is hyperexpressed in AITD glands are issues relevant to the above hypothesis. Results from immunofluorescence and Northern blotting studies on primary thyrocyte cultures and on a thyroid cell line demonstrate that thyrocytes express constitutively TAP-1 at a low level, and that this expression is readily induced by interferon-gamma (IFN-γ) and to a lesser extent by IFN-α. In AITD, but not in non-autoimmune glands, thyrocytes hyperexpress TAP-1, as demonstrated by both immunohistopathology and flow cytometry. The cytokine pattern does not bear, as assessed by reverse transcriptase-polymerase chain reaction (RT-PCR), a clear relationship with TAP-1 expression. These results have broad implications and suggest that the core concept of the ‘aberrant HLA expression’ hypothesis of endocrine autoimmunity could be incorporated in the currently prevailing view of ‘autoimmunity by breach of peripheral tolerance’.
Non-restricted T cell receptor (TCR)-Vα and -Vβ gene usage in patients with pulmonary sarcoidosis
Authors: HOSHINO T , SUZUKI R , TSURUTA Y , MATSUTANI T , KIKUCHI M , KAWAMOTO M , GOUHARA R , SHIRAISHI T , MOCHIZUKI M , ITOH K , OIZUMI K
Published: November 2003

Sarcoidosis is a systemic granulomatous disease of unknown etiology characterized by the pronounced accumulation of CD4+ T cells and macrophages in the affected organs. TCR variable (V)α and Vβ gene usage in patients with sarcoidosis is still a matter of discussion. In this investigation, we analysed TCR-Vα and -Vβ gene usage in bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells (PBMC) of 30 patients with active pulmonary sarcoidosis using an adapter ligation method, reverse transcriptase-polymerase chain reaction (RT-PCR), and sequence-specific oligonucleotide probe (SSOP) analyses. There was no significant difference in TCR-Vα or -Vβ gene usage between BALF (n=12) or PBMC (n=27) of patients and PBMC of healthy subjects (n=10). Neither selective TCR-Vα nor -Vβ expansion was observed in the paired BALF and PBMC from seven of nine patients. However, selective expansions were observed in a few TCR-Vα or -Vβ subsets in the BALF or PBMC of some individuals. Although a modest increase in a few TCR-Vα or -Vβ subsets was observed in the BALF or PBMC of some individuals, the increased TCR-Vα or -Vβ subsets were not closely associated with the HLA-DRB1, DQA1, DQB1, and DPB1 alleles of these patients. These results suggest that TCR-Vα or -Vβ gene usage is not restricted in both lung and peripheral blood in the majority of patients with active pulmonary sarcoidosis.
A non-protective T helper 1 response against the intra-macrophage protozoan Theileria annulata
Authors: CAMPBELL J D M , BROWN D J , NICHANI A K , HOWIE S E M , SPOONER R L , GLASS E J
Published: November 2003

Theileria annulata is a protozoan parasite which infects and transforms bovine macrophages. Infected macrophages possess augmented antigen presentation capabilities, as they are able to activate the majority of T cells from unexposed animals. In vivo, T cells in the draining lymph node (principal site of parasite development) are activated ‘non-specifically’ by the parasite. This event is followed by failure of the immune response to control the infection. Protective immune responses against intra-macrophage protozoa are usually mediated by T helper 1 (Th1) T cell responses. Here we examine the cytokine responses made by T. annulata-activated T cells. We show that the outcome of in vitro activation of T cells by parasitized macrophages is a skewing of their cytokine responses towards preferential expression of interferon-gamma (IFN-γ) mRNA. The in vitro response is mirrored during in vivo infection, as greatly elevated amounts of IFN-γ protein are found in lymph efferent from infected lymph nodes, while expression of IL-4 mRNA within the node stops. IFN-γ production does not correlate with protection against the parasite, as infected cells flourish during peak IFN-γ production, and only very small amounts of IFN-γ are produced during the effective immune response of an immunized animal. Overproduction of IFN-γ and loss of IL-4 expression are also likely to account for the failure of B cells to reach the light zone of germinal centres, a developmental step which is tightly regulated by cytokines.
Cellular mRNA expression of interferon-gamma (IFN-γ), IL-4 and IL-10 relates to resistance to experimental autoimmune myasthenia gravis (EAMG) in young Lewis rats
Authors: SHI F-D , ZHANG G-X , BAI X-F , VAN DER MEIDE P-H , LINK H
Published: November 2003

Age-related alterations in the immune system, including changes in lymphocyte subset composition, result in changes of cytokine patterns and might thereby influence the incidence and severity of autoimmune diseases. To investigate the age-related resistance to EAMG, an animal model for human MG, young (4-week-old) and adult (8–10-week-old) female Lewis rats were immunized with Torpedo acetylcholine receptor (AChR) and Freund's complete adjuvant (FCA). Adult Lewis rats showed severe weight loss and progressive muscular weakness after immunization, while young rats developed minor clinical signs of EAMG after a prolonged interval post-immunization. By comparison with adult rats, the young had lower AChR-specific T and B cells responses, and less muscle AChR loss. In situ hybridization performed on mononuclear cells (MNC) from lymph nodes revealed that young rats had lower levels of AChR-specific IFN-γ, IL-4 and IL-10 mRNA-expressing cells compared with adult rats. Since IFN-γ, IL-4 and IL-10 promote the development of EAMG, the low expression of these cytokines might contribute to EAMG resistance in young Lewis rats.
Effect of body weight and caloric restriction on serum complement proteins, including Factor D/adipsin: studies in anorexia nervosa and obesity
Authors: POMEROY C , MITCHELL J , ECKERT E , RAYMOND N , CROSBY R , DALMASSO A P
Published: November 2003

Complement plays important roles in host immune defences, and recent studies suggest that adipose tissue is an important site of production for some complement proteins. Starvation has been associated with low complement levels, but studied populations have usually had concomitant opportunistic infections or other conditions which might affect complement levels. To determine the impact of body weight and changes in body weight on serum complement, we investigated levels of complement proteins in otherwise healthy patients with a wide range of body weights, including patients with anorexia nervosa before and after treatment, obese dieters before and after weight loss, and normal weight controls. We found that complement proteins of the alternative pathway (C3, B, and D), alternative pathway haemolytic activity (AP50) and the inhibitors H and I were low in starving anorectics and normalized with weight gain. C3a levels were comparable in anorectics at low weight and after weight gain, indicating that low serum complement levels were attributable to hypoproduction and not complement cascade activation with consumption. Further, levels of C3, B, AP50, H and I, but not D, were higher than controls in obese patients and decreased toward normal after weight loss. Overall, percentage of ideal body weight, changes in body weight, and serum transferrin were each highly correlated with serum levels of complement proteins. We conclude that levels of alternative pathway complement components are determined in part by factors that influence body weight and by weight changes, possibly due to changes in production in adipose tissue or at other sites.
Amelioration of collagen II-induced arthritis in rats by the type IV phosphodiesterase inhibitor Rolipram
Authors: NYMAN U , MÜSSENER Å , LARSSON E , LORENTZEN J , KLARESKOG L
Published: November 2003

The effect of Rolipram, a selective inhibitor of the cyclic AMP specific phosphodiesterase (PDE IV) was evaluated in the rat collagen type II (RCII)-induced arthritis model in the DA rat. Rolipram was given either shortly before expected onset of disease (days 10–14) or shortly after the onset of clinically evident arthritis (days 15–19 after immunization). Administration at days 10–14 delayed the onset of arthritis for approximately 5 days, but the severity of arthritis was thereafter comparable to that seen in a non-treated control group. Rolipram treatment of animals with manifest arthritis inhibited further arthritis development and also tended to diminish its severity at a phase of disease where non-treated control animals showed a rapidly progressing disease development. Serum levels of antibodies to RCII were in all experiments similar between Rolipram-treated and control animals. An in situ hybridization method for determining cytokine mRNA synthesis in regional lymph nodes, after administration of Rolipram (at days 2–7), demonstrated a strong inhibitory effect on tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) mRNA expression, whereas no effects were seen on IL-2 mRNA synthesis after in vivo challenge with native RCII emulsified in Freund's incomplete adjuvant. The results thus demonstrate strong preventive as well as therapeutic effects of Rolipram in a model for arthritis that is very similar to human rheumatoid arthritis with respect to cytokine regulation, and suggest that Rolipram has its major effects in the effector stage of the arthritogenic immune response.
Restricted expression of FcγRIII (CD16) in synovium and dermis: implications for tissue targeting in rheumatoid arthritis (RA)
Authors: EDWARDS J C W , BLADES S , CAMBRIDGE G
Published: November 2003

Interactions between immune complexes and immunoglobulin Fc receptors may contribute to inflammation in RA. Previous studies suggested that FcγRIII (CD16) may be preferentially expressed in diseased synovial intima. The distribution of immunoreactive FcγRIII was examined in normal fetal limb tissues, and both normal and selected abnormal samples of adult synovium and skin. In fetal limbs at 10–14 weeks gestation FcγRIII was restricted to synovial intima. In normal adult synovium FcγRIII was restricted to intimal cells. In inflamed synovia differential expression of FcγRIII in the intima was less consistent. In both fetal and adult synovium FcγRIII was largely restricted to cells expressing CD45 (leucocyte common antigen). Staining for FcγRIII was, however, occasionally associated with CD45− intimal cells in fetal synovium. In both fetal and adult tissues cell membrane FcγRIII was frequently closely associated with complement decay-accelerating factor (DAF), which is present on intimal fibroblasts and extracellular matrix. FcγRIII expression was minimal in normal forearm dermis, but widespread on CD45+ cells in skin exposed to mechanical stress. In skin containing rheumatoid nodules, FcγRIII was preferentially expressed on palisading macrophages. These observations indicate that expression of FcγRIII on macrophages may be involved in the susceptibility of connective tissues to immune complex-induced damage in RA. Colocalization of FcγRIII and DAF in synovium may indicate an unrecognized functional interrelationship.
Mechanisms of T cell-induced glomerular injury in anti-glomeruler basement membrane (GBM) glomerulonephritis in rats
Authors: HUANG X R , TIPPING P G , APOSTOLOPOULOS J , OETTINGER C , D'SOUZA M , MILTON G , HOLDSWORTH S R
Published: November 2003

The effector mechanisms of T cell-dependent acute glomerular injury were studied in autologous phase anti-GBM glomerulonephritis (GN) in rats. Acute proliferative GN was induced in sensitized rats by a subnephritogenic dose of sheep anti-rat GBM antibody. Injury was manifested by proteinuria and glomerular leucocyte infiltration composed predominantly of macrophages but also CD4+ and CD8+ T cells. T cell depletion, using an anti-CD5 MoAb, demonstrated that glomerular leucocyte infiltration and proteinuria were T cell-dependent. Inhibition of T helper cell function using an anti-CD4 MoAb prevented proteinuria and glomerular macrophage and CD4+ T cell influx, but not accumulation of CD8+ T cells. Depletion of CD8+ T cells also prevented proteinuria and the influx of macrophages and CD8+ T cells, but not accumulation of CD4+ T cells. Macrophage depletion, using micro-encapsulated clodronate, prevented proteinuria and glomerular macrophage infiltration, but not the accumulation of CD4+ or CD8+ T cells, indicating that macrophages are the common cellular effectors for both CD4 and CD8 T cell-dependent injury. Evidence for cytotoxic mechanisms of injury (increased numbers of apoptotic cells or accumulation of natural killer (NK) cells in glomeruli) could not be demonstrated. These data suggest that acute glomerular injury in anti-GBM GN is the result of macrophage recruitment, which is dependent on both CD4 and CD8 T cells, and that direct T cell-mediated injury (cellular cytotoxicity) is not involved.
Persistent production of interferon-gamma (IFN-γ) and IL-12 is essential for the generation of protective immunity against Listeria monocytogenes
Authors: XIONG H , OHYA S , TANABE Y , MITSUYAMA M
Published: November 2003

IFN-γ and IL-12 are believed to be important in the host defence against Listeria infection in mice. However, the relationship between these two cytokines and generation of protective immunity remains poorly understood. In the present study, it was found that at least 4 days of immunizing infection were required for the generation of protective immunity against L. monocytogenes. Protective immunity was generated only by immunizing infection with virulent strain. Even repeated injections of avirulent strain failed to induce protective immunity. When the immunizing infection was terminated with antibiotics, generation of protective immunity and IFN-γ-producing ability was impaired, while expression of IFN-γ and IL-12 was also impaired. The mutual relationship between IFN-γ and IL-12 in L. monocytogenes infection was analysed in vitro. After neutralization of IL-12, IFN-γ production was completely blocked and IFN-γ expression was also inhibited. In contrast, there was no change of IL-12 expression after neutralization of IFN-γ. Taking all facts into consideration, it may be concluded that persistent production of IFN-γ induced by persistent production of IL-12 during immunizing infection is essential for the generation of protective immunity against L. monocytogenes.
Reduction of the DTH response is related to morphological changes of Langerhans cells in mice exposed to acute immobilization stress
Authors: KAWAGUCHI Y , OKADA T , KONISHI H , FUJINO M , ASAI J , ITO M
Published: November 2003

There is a large body of evidence indicating that stress influences immune competence. For example, psoriasis and atopic dermatitis may be exacerbated by psychic stress and related to abnormalities in the cellular constituents of the immune system in the skin. However, the underlying mechanisms remain unclear. We therefore investigated the potential of acute immobilization stress to affect the DTH response in BALB/c mice. DTH was significantly reduced in an immobilization time-dependent manner when stress exposure was just before sensitization. Although the number of Langerhans cells (LC) did not change under these conditions, marked alteration of LC morphology was observed with a significant decrease in area. Recovery of LC was observed within 24 h when the DTH response was also restored. Expression of the calcitonin gene-related peptide (CGRP), which inhibits LC antigen presentation, was significantly increased up to 1.6-fold in nerve fibres of immobilized mice. We conclude that stress-induced suppression of DTH could be due to reduction of LC antigen presentation with morphological change in association with CGRP elevation.
Humoral response against heat shock proteins and other mycobacterial antigens after intravesical treatment with bacille Calmette–Guérin (BCG) in patients with superficial bladder cancer
Authors: ZLOTTA A R , DROWART A , HUYGEN K , DE BRUYN J , SHEKARSARAI H , DECOCK M , PIRSON M , JURION F , PALFLIET K , DENIS O , MASCART F , SIMON J , SCHULMAN C C , VAN VOOREN J P
Published: November 2003

Few studies have analysed the antibody response during intravesical BCG immunotherapy for superficial bladder cancer. We have examined the evolution in serum antibody response against several heat shock proteins (hsp), including the recombinant mycobacterial hsp65 and the native protein P64 from BCG, GroEL from Escherichia coli (hsp60 family), recombinant mycobacterial hsp70 and the E. coli DnaK (hsp70 family), against purified protein derivative of tuberculin (PPD) and the AG85 complex of Mycobacterium bovis BCG, as well as against tetanus toxoid in 42 patients with a superficial bladder tumour, 28 treated with six intravesical BCG instillations and 14 patients used as controls. We also analysed the lymphoproliferative response of peripheral blood mononuclear cells against PPD in this population. Data of antibody responses at 6 weeks post BCG were available in all 28 patients, and at 4 month follow up in 17 patients. All patients who demonstrated a significant increase in IgG antibodies against PPD at 4 months follow up had a significant increase already at 6 weeks of follow up. In contrast, IgG antibodies against hsp increased significantly from 6 weeks to 4 months post-treatment. A significant increase in IgG antibodies against PPD, hsp65, P64, GroEL, and hsp70 at 4 months follow up was observed in 10/17, 8/17, 10/17, 4/17 and 8/17 patients. Native P64 protein elicited a higher antibody response than recombinant mycobacterial hsp65. No increase in antibody response was observed against Dnak from E. coli, against AG85 or tetanus toxoid after BCG therapy. An increase in IgG antibodies against P64 at 4 months follow up compared with pretreatment values was found to be a significant predictor of tumour recurrence (P < 0.01). Further studies with a larger number of patients are needed to confirm the value of the antibody response against P64 as a clinical independent prognostic factor.
Rescue by cytokines of apoptotic cell death induced by IL-2 deprivation of human antigen-specific T cell clones
Authors: KANEKO S , SUZUKI N , KOIZUMI H , YAMAMOTO S , SAKANE T
Published: November 2003

The control of cell survival and cell death is of central importance in tissues with high cell turnover such as the lymphoid system. We have examined the effect of cytokines on IL-2 deprivation-induced apoptosis of human antigen-specific T helper clones with different cytokine production profiles. We found that IL-2, interferon-alpha (IFN-α), and IFN-β inhibited IL-2 deprivation apoptosis in Th0, Th1, and Th2 clones. We also found that IL-2 protects T cell clones from IL-2 deprivation apoptosis accompanying active proliferation and enhanced expression of P53, Rb and Bcl-xL proteins. In contrast, IFN-α/β rescued T cell clones from apoptosis without active proliferation, and expression of apoptosis-associated proteins tested so far was unaffected. This may be due to the fact that T cells treated with IL-2 contained those located in S+G2/M phases of the cell cycle, whereas the vast majority of T cells treated with IFN-α/β were located in G0/G1 phase. IFN-α/β specifically induced tyrosine phosphorylation and translocation into nucleus of signal transducers and activators of transcription (STAT) 2 protein in the T cell clones. In addition, over-expression of STAT2 by transfection of the cDNA prevented apoptosis of the T cell clones. Our present study shows that IFN-α and -β mediate anti-apoptotic effect through other pathways than that of IL-2 in growth factor deprivation apoptosis.
Growth of recombinant Mycobacterium tuberculosis H37Ra in mouse macrophages
Authors: FALCONE V , COLLINS F
Published: November 2003

Mycobacterium tuberculosis H37Rv and H37Ra were derived from the same parental strain but differ strikingly in their virulence for experimental animals. Transfer of genetic material between these closely related strains resulted in the isolation of a number of recombinant H37Ra clones bearing the in vivo growth-promoting ivg locus of H37Rv. The recombinant strain was phagocytosed by murine peritoneal macrophages infected in vivo or in vitro and their intracellular growth rates were compared with the vector control. The intracellular growth of the recombinant was significantly faster than the vector control, but substantially slower than the wild-type H37Rv control, regardless of the method used to infect the macrophages. The slower intracellular growth observed for the recombinant strains was not due to a genetically induced metabolic defect, since they grew in synthetic liquid medium at rates equal to those observed for both H37Rv and H37Ra. Peritoneal macrophage monolayers provide a rapid and convenient assay by which to screen H37Ra recombinants for the presence of putative virulence genes.
Intracerebral injection of myelin basic protein (MBP) induces inflammation in brain and causes paraplegia in MBP-sensitized B6 mice
Authors: TSAI C Y , CHOW N H , HO T S , LEI H Y
Published: November 2003

Brain inflammation and paraplegia can be induced by an additional intraperitoneal (i.p.) and intracerebral (i.c.) restimulation in B6 mice after standard immunization with MBP in Freund's complete adjuvant (FCA) and Bordetella pertussis coadjuvant. Only the combination of i.p. MBP/FCA and i.c. MBP injection could induce clinical paraplegia; either one alone was not effective. Clinical symptoms would develop 2 days after the i.c. injection. The induction of paraplegia was MBP-specific, as irrelevant bovine serum albumin with the same protocol could not induce it. The i.p. restimulation was requisite and needed the MBP in FCA, as MBP in PBS was ineffective. Histopathological observation manifested cellular infiltration by leucocytes in perivascular spaces and cerebral cortex. Neutrophils were prominent at 12 h after i.c. injection, then were replaced by mononuclear cells 24 h later. There were dynamic changes in cell number and immunophenotype of VLA-4+ expression in cervical lymph node cells after i.c. injection. The cells derived from cervical lymph nodes had higher MBP-stimulated proliferation than that of distal lymph nodes. This additional i.p. and i.c. stimulation provides a new manipulation to study brain inflammation.
Naive and memory T cell infiltrates in chronic hepatitis C: phenotypic changes with interferon treatment
Authors: IMADA K , FUKUDA Y , KOYAMA Y , NAKANO I , YAMADA M , KATANO Y , HAYAKAWA T
Published: November 2003

The phenotypes of infiltrating lymphocytes in liver with chronic hepatitis C, including changes associated with interferon (IFN) treatment, were characterized. Specimens obtained from 22 patients treated with IFN were examined using avidin-biotin-peroxidase immunohistochemistry. In areas of lobular and periportal inflammation, most lymphocytes were CD8+ T cells of the CD45RO+ (memory) subset. The centres of lymphoid follicles were occupied by CD20+ B cells and a few CD4+ T cells which were CD45RA+ (naive subset). Follicular centres were surrounded mainly with CD4+ T cells. CD8+ T cells, mostly CD45RO+, were scattered through the mantle zones of follicles and extended around them. No significant changes in CD45RA+ lobular infiltrates accompanied IFN treatment. On the other hand, the number of CD45RO+ lobular infiltrates decreased after IFN treatment in complete responders (P <0.01). Moreover, there were significant correlations between CD45RO+ cell counts and serum alanine aminotransferase concentrations, CD45RO+ cell counts and the liver histologic grade and CD45RO+ cell counts and CD8+ cell counts. These results suggest that CD8+ memory T cells participate in hepatocyte injury in chronic hepatitis C, and that a decrease of CD8+ memory T cells correlates with the decreased liver inflammation with IFN treatment.
Induction and suppression of anti-antibodies to syngeneic T cell-binding antibodies in mice
Authors: KREMMER E , MYSLIWIETZ J , THIERFELDER S
Published: November 2003

Considerable effort is being invested in antibody gene technologies for production of species-adapted anti-T cell antibodies which can overcome formation of neutralizing anti-antibodies (anti-Ab). By establishing a mouse model for the generation of syngeneic anti-T cell MoAb, we addressed the question of whether ideally species-adapted T cell-binding antibodies can prime mice to produce anti-Ab. Two anti-Thy-1.2 MoAbs of IgG2a (MmTC) or IgM (MmTC-IgM) isotype were generated in congenic C57Bl/6-Thy-1.1 mice. Three injections of MmTC in C57Bl/6-Thy-1.2 or (C57Bl/6-Thy-1.2×C57Bl/6-Thy-1.1)F1 hybrids where they act as syngeneic anti-T cell MoAbs induced low anti-Ab. When MmTC was injected as Igh-mismatched antibody in CBA/J mice, high titre anti-MmTC antibodies were measured and fully mismatched skin allografts rejected within 26 days. MmTC injected twice weekly in syngeneic C57Bl/6 mice induced low anti-MmTC and prolonged graft survival up to day 117. In contrast, retreatment induced anti-MmTC with graft rejection within 32 days. Thus, upon retreatment, the immunosuppressive effects of MmTC were inhibited to a similar extent as that seen after antibody treatment in Igh-mismatched mice. However, a recently analysed immunological principle to suppress anti-Ab against T cell binding allo- or xenoantibodies by preinjection of T cell-depleting antibody with species differences in heavy chains also suppressed syngeneic anti-Ab after MmTC retreatment. This effect was accompanied by prolonged graft survival. Thus, our data indicate that inhibitory anti-Ab must be considered in humans even when treated with fully species-adapted anti-T cell MoAb, but may be suppressed by preinjection of a Fc region-mismatched anti-T cell antibody.
Cellular requirements for immunomodulatory effects caused by cell wall components of Paracoccidioides brasiliensis on antibody production
Authors: SILVA M F , BOCCA A L , FERRACINI R , FIGUEIREDO F , SILVA C L
Published: November 2003

In a previous study, we reported an increase in the number of immunoglobulin-secreting cells and the augmentation of antibody production (IgM and IgG3) against unrelated antigens (sheep erythrocytes or bovine serum albumin (BSA)) in mice infected with the fungus Paracoccidioides brasiliensis as well as in mice inoculated with its cell wall preparation (CW). The immunomodulatory effect of the live fungus and CW preparation was dose-dependent and mainly restricted to the i.p. inoculation simultaneously to the BSA challenge by the i.v. route. In the present study, we investigated the active component of CW preparation upon the phenotype and also the degree of activation of possible target peritoneal cells involved in those phenomena. An insoluble polysaccharide fraction (F1 fraction) mainly composed of β-glucan and chitin, and the purified β-glucan (BGPb) behaved as CW in the augmentation of early antibody production. The peritoneal mononuclear inflammatory cells induced by CW, F1 fraction and BGPb were highly positive to α-naphthyl esterase staining; released low H2O2; expressed high levels of MHC-Iad molecules and produced inflammatory cytokines such as tumour necrosis factor-alpha (TNF-α) and IL-6. Phenotypic analysis by flow cytometry and immunohistochemical techniques of the inflammatory cells responding to F1 fraction showed a prevalence of (CD11b/CD18, Mac-1)+ peritoneal macrophages. In addition, s.c. inoculation of F1 fraction resulted in the formation of nodular, localized and not progressive granulomatous lesions with an accumulation of (CD11b/C18)+ macrophages. Adoptive transferred Mac-1 macrophages to immunized syngeneic recipient mice were able to cause an increase in anti-BSA antibody production. These results suggest that inflammatory (CD11b/CD18)+ macrophages may be related to immunological disturbances, caused by cell wall components of P. brasiliensis.