bluuebunny/crossref_metadata_2025 huggingface dataset:
Induction of Apoptosis and Cytokine Gene Expression in T‐Cell Lines by Sera of Patients with Systemic Lupus Erythematosus
Authors: YANG B.‐C. , WANG Y.‐S. , LIN L.‐C. , LIU M.‐F.
Published: November 2003

Accelerated apoptosis and improper expression of cytokine genes have been considered as important defects of lymphocytes for the development of systemic lupus erythematosus (SLE). This study was undertaken to test the possible contribution of serum factors obtained from SLE patients to these abnormalities. Molt‐4 and Jurkat cells constantly exhibited a slower growth rate as well as more dead cells in culture with SLE sera tested than controls, although the cell cycle progression was apparently unaffected. Increased apoptosis was demonstrable among SLE sera‐cultured cells by ELISA for apoptosis‐specific DNA fragments and terminal deoxynucleotidyl transferase (TdT) in situ death analysis. Different levels of Fas, Fas‐L, and Bcl‐2 gene products were not detected between SLE sera‐treated cells and the controls. The transcripts of interleukin‐10 (IL‐10) and interferon‐γ (IFN‐γ) genes of these two T cell lines were evidently increased in the presence of SLE sera, while IL‐2 and IL‐4 were unaffected. Elevated expression of IL‐5 was also found in Molt‐4 cells. By contrast, SLE sera reduced the transcripts of IL‐6 gene in Jurkat cells. The effects of SLE sera were independent of corticosteroid medication. These results suggest that serum abnormalities may also play a role in T cell dysfunction.
IgG2b Inducing Factor from Rheumatoid Arthritis Synovial Fluid Synergizes with Transforming Growth Factor‐β in Promoting IgG2b Antibody Production in Mouse B Lymphocytes
Authors: LETTESJO¨ H. , RIDDERSTAD A. , MO¨LLER E.
Published: November 2003

Rheumatoid arthritis synovial fluid (RA‐SF) contains a distinct biological activity that selectively induces IgG2b production in LPS‐activated murine B blasts. This IgG2b inducing factor (IgG2bIF) acts directly on purified LPS‐activated B blasts from normal and Bruton's tyrosine kinase‐defective CBA/N mice. In order to test the possibility that TGF‐β and IgG2bIF in RA‐SF act in concert to induce IgG2b production, anti‐TGF‐β monoclonal antibodies and RA‐SF were added to LPS‐activated CBA B blasts, which led to a marked reduction of IgG2b‐producing cells. This result indicates that TGF‐β and IgG2bIF in RA‐SF synergize in the induction of IgG2b production. TGF‐β antibodies do not inhibit IgG2b production in CBA/N B blasts, further substantiating our earlier notion that CBA/N B blasts have a higher endogenous production of TGF‐β after LPS stimulation, which might be responsible for the aberrant reactivity in btk deficient CBA/N mice. RA‐SF is able to reconstitute the deficient IgG1 response in LPS‐ and IL‐4‐stimulated CBA/N B blasts. Addition of antibodies against TGF‐β had only a marginal effect on the IgG1 response, indicating that the reconstitution is mediated by another factor(s), which is present in RA‐SF.
IgG2b Inducing Factor from Rheumatoid Arthritis Synovial Fluid Activates Antibody Production in Highly Purified Mouse B Lymphocytes
Authors: LETTESJO¨ H. , MO¨LLER E.
Published: November 2003

Rheumatoid arthritis synovial fluid (RA‐SF) contains a factor that induces IgG2b antibody production in LPS‐stimulated murine B cells and therefore is called IgG2b inducing factor (IgG2bIF). When LPS, together with crude RA‐SF or semi‐purified IgG2bIF, was added to highly purified LPS‐stimulated B cells, the number of IgG2b‐producing cells was substantially enhanced. This shows that IgG2bIF acts directly on activated B cells, presumably by binding to a receptor expressed on LPS‐activated B cells. In vivo LPS‐activated B blasts were not able to respond to RA‐SF unless LPS was present in vitro, showing that LPS is needed to maintain cell viability and responsiveness to the IgG2bIF. To elucidate the mechanism for the IgG2bIF effect on highly purified B cells, the IgG2b response of LPS‐stimulated, Bruton's tyrosine kinase‐defective, xidB blasts was studied. Purified B blasts from the btk‐defective CBA/N mouse strain were sensitive to IgG2bIF. These findings show that IgG2bIF acts directly on B cells and activates cells through a btk‐independent pathway.
Predominance of Detrimental Humoral Immune Responses to HIV‐1 in AIDS Patients with CD4 Lymphocyte Counts Less Than 400/mm3
Authors: MCDOUGALL B. , NYMARK M. H. , LANDUCCI G. , FORTHAL D. , ROBINSON JR. W. E.
Published: November 2003

The humoral immune response to human immunodeficiency virus type 1 (HIV‐1) was studied in 25 AIDS patients with CD4 lymphocyte counts of less than 400/mm3. Humoral immune responses against tissue culture adapted strains of HIV‐1, and two limited‐passage patient isolates were investigated. Total anti‐HIV antibody levels were not significantly different between different individuals. Neutralizing titres against HIVLAI and HIVSF2 were 10‐ to 100‐fold higher than against clinical isolates. The complement‐mediated, antibody‐dependent enhancement of HIV‐1 infection titre was high (mean 1:14,000). Antibody‐complement mediated cytotoxicity of both HIVLAI and HIVSF2 was ineffective using human complement as a complement source. The antibody‐dependent, cell‐mediated cytotoxicity (ADCC) activity varied against the four isolates with tissue culture‐adapted strains being more susceptible than clinical isolates. Finally, an ADCC effector cell function, natural killer or NK activity, was measured for all 25 patients, and NK activity of patients was decreased by nearly 75% compared to uninfected individuals. In summary, beneficial humoral immune responses are low in HIV‐1 infected individuals with CD4 counts of less than 400/mm3 if the in vitro assay system is constructed to best mimic the in vivo situation. These results suggest that the lack of functional antibody responses to HIV may play an important role in viral pathogenesis.
Detection of Alterations in the Levels of Neuropeptides and Salivary Gland Responses in the Non‐Obese Diabetic Mouse Model for Autoimmune Sialoadenitis
Authors: YAMAMOTO H. , ISHIBASHI K. , NAKAGAWA Y. , MAEDA N. , ZENG T. , ROBINSON C. P. , OXFORD G. E. , CHEGINI N. , HUMPHREYS‐BEHER M. G.
Published: November 2003

The salivary glands of non‐obese diabetic (NOD) mice and BALB/c controls were evaluated for the stimulatory effects of the following neuropeptides; substance P (SP), vasoactive intestinal polypeptide (VIP), and neuropeptide Y (NPY). Injection of either of the three neuropeptides in combination with the muscarinic–cholinergic agonist pilocarpine increased saliva flow rates in BALB/c mice while there was no observable augmentation to flow rates in pre‐diabetic or diabetic NOD mice. Small increases in protein content of the stimulated saliva were observed in the BALB/c group of animals with the injection of any of the above neuropeptides in combination with pilocarpine. In pre‐diabetic NOD animals, only VIP and NPY increased the protein content‐ratio above pilocarpine alone. Radioimmunoassay determination of neuropeptide concentrations in the submandibular and parotid glands revealed reduced levels of SP with diabetes onset as compared with pre‐diabetic NOD or BALB/c mice. The levels of NPY were similar between BALB/c and NOD animals except in the pre‐diabetic parotid gland where NPY concentrations were 1.3‐fold greater. On the other hand, VIP concentrations were substantially reduced in the submandibular gland of NOD mice, while in the parotid gland neuropeptide levels were evaluated 3.8‐fold relative to BALB/c controls. Immunohistochemical staining of the parotid and submandibular glands for SP revealed primarily ductal cell staining which was reduced with diabetes onset in NOD animals. These findings further define the sialoadenitis observed in NOD mice to be due, in part, to a general loss of neurotransmitter responsiveness on the part of salivary gland cells.
Endogenous Adenosine Curtails Lipopolysaccharide‐Stimulated Tumour Necrosis Factor Synthesis
Authors: EIGLER A. , GRETEN T. F. , SINHA B. , HASLBERGER C. , SULLIVAN G. W. , ENDRES S.
Published: November 2003

Recent studies have demonstrated the inhibitory effect of exogenous adenosine on TNF production. During inflammation endogenous adenosine levels are elevated and may be one of several anti‐inflammatory mediators that reduce TNF synthesis. In the present study the authors investigated this role of adenosine in freshly isolated human PBMC. The effect of endogenous adenosine on TNF formation was studied by four different approaches. First, adenosine deaminase was added to LPS‐stimulated mononuclear cells. This enzyme specifically deaminates extracellular adenosine to the inactive metabolite inosine. TNF production was augmented from baseline stimulation (LPS alone) of 3.5 ± 0.4 ng ml−1— 5.2 ± 0.9 ng ml−1 in the presence of 10 U ml−1 adenosine deaminase. Second, TNF production was determined after stimulation in the presence of dipyridamole, an inhibitor of cellular re‐uptake of adenosine which increases extracellular concentrations. TNF synthesis was reduced dose‐dependently from 3.1 ± 0.9 ng ml−1— 1.1 ± 0.2 ng ml−1 by 10 μm dipyridamole. Third, the adenosine A2 receptor antagonist 8‐(3‐chlorostyryl)caffeine (100 nm) enhanced TNF synthesis from a baseline of 3.7 ± 0.5 ng ml−1— 5.5 ± 0.9 ng ml−1. In contrast, no increase resulted from the addition of 100 nm of the specific A1 receptor antagonist 8‐cyclopentyl‐1,3‐dipropylxanthine. Finally, the authors were able to show that suppression of TNF formation by the specific type IV phosphodiesterase inhibitor rolipram can be completely reversed by adenosine deaminase or by the application of the A2 receptor antagonist. The authors conclude that endogenous adenosine controls TNF production. This effect of adenosine may not only have a physiological role but also appears to contribute to the pharmacological inhibition of TNF synthesis by exogenous agents such as the specific type IV phosphodiesterase inhibitor rolipram.
The Stability of Lymphocytic β1,4‐Galactosyltransferase Expression During Pregnancy and Lactation
Authors: JEDDI P. A. , KEUSCH J. , LYDYARD P. M. , DELVES P. J.
Published: November 2003

The authors used murine pregnancy as a model to investigate the regulation of IgG glycosylation. Pregnancy is associated with decreased levels of circulating IgG. The oligosaccharides on this IgG from late (day 15), but not early (day 8), pregnant Balb/c mice exhibited increased levels of terminal galactose. The levels remained elevated 8 days post‐partum in lactating mice. Nonetheless, splenic β1,4‐GalTase mRNA and enzyme activity remained relatively constant throughout pregnancy and into lactation. This was in contrast to a pregnancy‐associated increase in mammary gland β1,4‐GalTase mRNA. Thus the increased IgG galactose levels seen in pregnancy are regulated by mechanisms which are independent of transcriptional control of β1,4‐GalTase expression.
Host Responses and Antigens Involved in Protective Immunity to Mycobacterium tuberculosis
Authors: ANDERSEN P.
Published: November 2003

Tuberculosis (TB) is the largest single infectious cause of human mortality. The incidence of TB has remained high in most of the developing world and the disease has recently re‐emerged as a public health problem in industrialized countries. The development of a new improved TB vaccine is a highly prioritized international research area, which has been further stimulated by the appearance of multi‐drug resistant strains of Mycobacterium tuberculosis. The present status of the attempts to characterize the protective immune response to TB will be reviewed with special emphasis on recent progress in the identification and characterization of target molecules recognized by protective cells. This paper will focus on proteins released from live bacteria and discuss their role in the host–pathogen interaction and the ongoing attempts to use these molecules in TB subunit vaccines.
IL‐4 Synergizes with IL‐10 and Anti‐CD40 MoAbs to Induce B‐Cell Differentiation in Patients with Common Variable Immunodeficiency
Authors: PUNNONEN J. , KAINULAINEN L. , RUUSKANEN O. , NIKOSKELAINEN J. , ARVILOMMI H.
Published: November 2003

In the present study the phenotype and function of lymphocytes from patients with common variable immunodeficiency (CVI) were studied. Five out of 12 patients had abnormally low proportion of CD4+ T cells, but PBMC of these patients were capable of proliferating in response to polyclonal T‐cell mitogens or PPD antigen. The phenotype of patients' B cells, as determined by expression of CD10, CD19 and CD34, was comparable to that of healthy controls. IL‐4 and anti‐CD40 MoAbs induced moderate B‐cell differentiation in PBMC derived from patients with CVI, but the frequencies of Ig‐secreting cells were generally at levels spontaneously observed in healthy controls. IL‐10 was completely ineffective in inducing IgG‐secreting cells in cultures of PBMC derived from patients with CVI even in the presence of anti‐CD40 MoAbs, whereas high frequencies of Ig‐secreting cells were induced under similar condition in cultures of PBMC derived from healthy controls. Importantly, when IL‐4 was added to cultures stimulated with anti‐CD40 MoAbs and IL‐10, a very strong synergistic effect on the numbers of Ig‐secreting cells and the levels of Ig secretion was observed in PBMC from both patients and controls. Moreover, the frequencies of Ig‐secreting cells after activation with anti‐CD40 MoAbs, IL‐4 plus IL‐10 in PBMC from some patients were comparable to those observed in PBMC from healthy controls. Taken together, these results indicate that B cells from patients with CVI have impaired capacity to differentiate into Ig‐secreting cells in response to IL‐10 and anti‐CD40 MoAbs, and that this unresponsiveness can be restored by exogenous IL‐4 in a proportion of the patients.
CD44 Expression by Leucocytes in Rheumatoid Arthritis and Modulation by Specific Antibody: Implications for Lymphocyte Adhesion to Endothelial Cells and Synoviocytes In Vitro
Authors: BRENNAN F.R. , MIKECZ K. , GLANT T.T. , JOBANPUTRA P. , PINDER S. , BAVINGTON C. , MORRISON P. , NUKI G.
Published: November 2003

Anti‐CD44 MoAb IM7 induced the loss of CD44 from mouse leucocytes thereby inhibiting leucocyte migration and joint inflammation in murine arthritis. Thus, targeting CD44 with MoAb may have potential for the treatment of patients with inflammatory joint diseases. Expression of CD44 by peripheral blood (PB) and synovial fluid (SF) leucocytes from rheumatoid arthritis (RA) patients was compared and the ability of IM7 to modulate this expression determined. RASF lymphocytes showed increased CD44 expression compared with those in PB indicative of an activated phenotype. As inflammatory SF did not up‐regulate CD44 expression on PB lymphocytes, the increased CD44 expression by SF lymphocytes was a result of the selective homing of CD44high cells to the synovium rather than an effect of the synovial environment. RASF granulocytes showed reduced CD44 expression compared with those in PB, again indicative of an activated phenotype. However, this reduction could be induced on PB granulocytes following culture with inflammatory SF and was inhibited by anti‐TNF‐α MoAb, implying that soluble factors in inflammatory SF such as TNF‐α induced granulocyte activation and CD44 loss. IM7 induced the loss of CD44 from lymphocytes (both from PB and SF) and granulocytes in vitro, but was subsequently re‐expressed after 24 h culture in the absence of the MoAb. This loss of CD44 was blocked by serine‐ and metalloprotease inhibitors implying that IM7 induced the proteolytic cleavage of CD44 by a mechanism similar to that reported for the loss of CD44 from PMA‐activated granulocytes. Furthermore, IM7‐treated CD44low lymphocytes showed reduced adherence to both an endothelial cell line and RA synovial fibroblasts in vitro. The unique ability of IM7 to reduce CD44 expression by lymphocytes suggests that it could prevent lymphocyte extravasation and synovial infiltration in RA as previously reported in murine arthritis.
Distribution of Monoclonal Antibodies in Intestinal and Urogenital Secretions of Mice Bearing Hybridoma ‘Backpack’ Tumours
Authors: HANEBERG B. , KENDALL D. , APTER F. M. , NEUTRA M. R.
Published: November 2003

Mice bearing IgA hybridoma ‘backpack’ tumours have been used to demonstrate that secretion of a single monoclonal IgA can protect against mucosal infection, but the relevance of this model to normal IgA protection is not clear. The authors analysed the distribution of specific monoclonal and total antibodies in bile, local intestinal secretions, cervical‐vaginal secretions, urine and serum of mice bearing anti‐cholera toxin (CT) IgA and IgG backpack tumours, with and without bile duct ligation. Backpack tumours resulted in high levels of both anti‐CT and total IgA or IgG in serum, and IgA (but not IgG) in bile. Secretions recovered by absorbent filter ‘wicks’ from mucosal surfaces throughout the intestines of backpack tumour mice contained significant concentrations of monoclonal anti‐CT IgA, but total IgA levels were as in normal mice. Neither monoclonal nor total IgA levels on mucosal surfaces were altered by bile duct ligation. Furthermore, anti‐CT monoclonal IgA levels in local intestinal secretions of backpack tumour mice were comparable to specific polyclonal IgA levels previously elicited by mucosal immunization with CT. Thus, IgA‐mediated protection against enteric challenge in the backpack tumour model may be a valid predictor of protection provided by natural mucosal immunization in vivo. High monoclonal IgA levels in bile, urine and the female genital tract, however, may not reflect the situation in normal immunized mice.
Interleukin‐7 (IL‐7) in Colorectal Cancer: IL‐7 is Produced by Tissues from Colorectal Cancer and Promotes Preferential Expansion of Tumour Infiltrating Lymphocytes
Authors: MAEURER M. J. , WALTER W. , MARTIN D. , ZITVOGEL L. , ELDER E. , STORKUS W. , LOTZE M. T.
Published: November 2003

Despite increasing survival rates for patients with colorectal cancer, additional treatment options are required, including active or passive immunotherapy for patients with metastatic disease. Freshly harvested colorectal cancer specimens and in vitro cultured colorectal cancer cell lines were examined for IL–7 protein secretion in order to examine the potential role of this cytokine in the interaction between tumour cells and the host immune system. Freshly harvested colorectal cancer specimens (21/21), or normal adjacent mucosa (3/3), as well as long‐term established colorectal cancer cell lines (3/4) exhibited IL‐7 mRNA expression as detected by RT‐PCR and confirmed by Southern Blot analysis. Freshly harvested colorectal cancer tissue (16/18), or long‐term established colorectal cancer cell lines (2/4) secreted in vitro IL‐7 as detected by ELISA. In contrast, breast, pancreatic, or lung cancer cell lines, as well as several haematopoietic cancer cells lines, tested negative for IL‐7 mRNA and protein. The authors tested different cytokines (IL‐1β, IL‐2, IL‐7, or a combination of IL‐1β/IL‐7) in vitro for the ability to expand tumour ‐ infiltrating T lymphocytes (TIL) from individual patients (n=9) with colorectal cancer. TIL populations were tested at day 14 after in vitro propagation for phenotypic analysis by FACS and for reactivity directed against NK and LAK sensitive target cells and autologous cancer cells as measured by cytotoxicity and cytokine release. TIL obtained from colorectal cancer lesions can be efficiently expanded in the presence of IL‐7, some (3/9) of which appear to exhibit autologous tumour recognition as measured by cytolytic effector functions and by detection of IFNγ and TNFα release. Detection of IL‐7 mRNA expression in colorectal cancer, in normal mucosa adjacent to tumour, as well as the ability of colorectal cancer tissue to secrete IL‐7, raises new questions about the biology of the host / tumour interactions in colorectal cancer.
Structure of the Mouse Serum Amyloid A 5 (Saa5) Gene: Relationship to Other Members of the Serum Amyloid A Family
Authors: BUTLER A. , WHITEHEAD A. S.
Published: November 2003

The mouse and human genes which encode the serum amyloid A (SAA) proteins have been extensively characterized. In the present study, the authors report the sequencing of the constitutively expressed mouse Saa5 gene, including ≈2 kilobases of promoter sequence which extends into the sequence of the adjacent Saa4 pseudogene. The Saa5 gene was thus determined to have a 4 exon/3 intron structure, with a B2 repeat located ≈450 base pairs upstream, and a microsatellite contained within intron 2. Analyses of Saa5 intronic and promoter sequences, by comparison with representative members of the mouse Saa and human SAA families, provided evidence which strongly suggests that mouse Saa5 and the constitutively expressed human SAA4 gene are evolutionary and functional homologues. Furthermore, putative transcription motifs were identified within the Saa5 and SAA4 promoters which may account, in part, for the constitutive expression and unique induction profiles of these genes.
Peyer's Patches in Murine AIDS: Dissociation Between Lymphoproliferation and Anergy
Authors: COLOMBI S. , MOUTSCHEN M. , DE LEVAL L. , DEMOITIÉ M.‐A. , TREBAK M. , MARTIN M.‐T. , BONIVER J.
Published: November 2003

RadLV‐Rs infection induces a murine immunodeficiency syndrome associated with a dramatic enlargement of spleen and lymph nodes. Surprisingly, the lymphoproliferation excludes thymus and Peyer's patches (PP). To understand the cellular interactions underlying lymphoproliferation further, the authors investigated the fate of PP in RadLV‐Rs infected mice. The atrophy of PP was mostly due to the depletion of B cells, while the proportion of CD4+ and CD8+ T cells was increased. Nevertheless, B cell phenotype was modified with the emergence of lymphocytes with a low expression of B220 in infected PP. T cells characterized by a memory/activated phenotype in control PP did not undergo phenotypical changes after viral infection (i.e. regarding Thy‐1 and CD44 expression). Despite the absence of lymphoproliferation, PP T and B cells displayed altered responses to mitogens in vitro. Finally, alterations of the expression of adhesion molecules and vascular addressins could not explain the atrophy of PP by a reduced homing to this lymphoid site. B cells and T cells from normal PP are clearly different from lymph nodes (LN) lymphocytes. The authors propose that the particular functional state which characterizes PP lymphocytes influences the B cell/T cell crosstalk necessary for RadLV‐Rs‐induced lymphoproliferation.
ICAM‐2 Provides a Costimulatory Signal for T Cell Stimulation by Allogeneic Class II MHC
Authors: CARPENITO C. , PYSZNIAK A. M. , TAKEI F.
Published: November 2003

To examine the functional role of intercellular adhesion molecule (ICAM)‐2 (CD102) in antigen presentation to T cells, the I‐Ed‐transfected murine fibroblastic L cell line RT10.3 (H‐2k) was transfected with murine ICAM‐1 or ICAM‐2 and tested for their abilities to stimulate C3H/He (H‐2k) splenic T cells. The expression of ICAM‐1 or ICAM‐2 on RT10.3 cells significantly increased the stimulation of T cells in an LFA‐1 (CD11a/CD18)‐dependent manner as determined by thymidine incorporation. This enhanced T cell response was also observed when combinations of untransfected RT10.3 cells and ICAM‐1‐ or ICAM‐2‐transfected L cells were used as stimulators, indicating that ICAM‐1 and ICAM‐2 deliver a costimulatory signal instead of merely enhanced T cell adhesion to antigen presenting cells. The T cells stimulated with ICAM‐transfected RT10.3 in the primary response vigorously responded to BALB/c (H‐2d) spleen cells in a secondary allogeneic stimulation. In contrast, T cells stimulated with untransfected RT10.3 in the primary response did not respond to BALB/c spleen cells in the secondary response. Significant secondary responses to a third party stimulator, C57BL/6 spleen cells (H‐2b), were observed regardless of ICAM expression on RT10.3 cells in the primary stimulation. These results indicate that ICAM‐2 as well as ICAM‐1 not only enhance antigen presentation mediated by allogeneic class II MHC but also provide a costimulatory signal to T cells. This costimulatory signal may be important in the aversion of an anergic state.
Serum Level of Soluble Interleukin‐2 Receptor in Patients with Seasonal Allergic Rhinitis
Authors: OHASHI Y. , TANAKA A. , KAKINOKI Y. , OHNO Y. , SAKAMOTO H. , KATO A. , MASAMOTO T. , WASHIO Y. , NAKAI Y.
Published: November 2003

The rate of release of the soluble form of interleukin‐2 receptor (soluble IL‐2R) reflects T cell activation in vivo. Since T lymphocytes play a central role in respiratory allergic disorders, the measurement of serum levels of soluble IL‐2R may be useful in analysing the disease state of allergic disorders. The study has aimed at investigating the seasonal changes in serum soluble IL‐2R in 81 patients with seasonal allergic rhinitis due to Japanese cedar pollens, with special reference to the effect of anti‐allergic pharmacotherapy and immunotherapy. Serum samples were obtained twice from each patient, before and during the pollen season, and all the serum samples were simultaneously used for determination of soluble IL‐2R and cedar pollen‐specific immunoglobin E (IgE). Seasonal elevation in soluble IL‐2R was not associated with the good clinical outcome but was associated with the poor clinical outcome, irrespective of pharmacotherapy or immunotherapy. Additionally, successful immunotherapy suppressed seasonal elevation of serum soluble IL‐2R more strongly than successful pharmacotherapy, and seasonal increase rates in soluble IL‐2R were inversely correlated with the duration of immunotherapy. Seasonal increase rates in soluble IL‐2R were significantly correlated with seasonal increase rates in specific IgE in both the medication group and the immunotherapy patients. These results may suggest that seasonal changes in serum soluble IL‐2R may serve as an indicator for clinical outcome of seasonal allergic rhinitis, that the magnitude of T cell activation could affect specific IgE production, and that T cell activation could be gradually modulated as immunotherapy proceeds. In conclusion, seasonal changes in serum soluble IL‐2R may serve as an objective indicator for clinical outcome of seasonal allergic rhinitis.
Interleukin‐10 Down‐Regulates Oxidative Metabolism and Antibody‐Dependent Cellular Cytotoxicity of Human Neutrophils
Authors: CAPSONI F. , MINONZIO F. , ONGARI A. M. , CARBONELLI V. , GALLI A. , ZANUSSI C.
Published: November 2003

The authors investigated the ability of interleukin‐10 (IL‐10) to modulate some constitutive or interferon‐γ (IFN‐γ)‐enhanced activities of human neutrophils. An 18 h culture of neutrophils with IL‐10 dose‐dependently down‐regulated their capacity to produce O2− and lucigenin‐amplified chemiluminescence in response to n‐formyl‐methionyl‐leucylphenyl‐alanine (FMLP). Furthermore, treatment of neutrophils with IL‐10 decreased in a dose‐dependent fashion, their capacity to lyse antibody‐coated sheep erythrocytes. Membrane expression of FcγRI, FcγRII, FcγRIII, CR1, CR3 and FcγR‐ and CR‐mediated phagocytosis were not modified by the cytokine. Culture of neutrophils with IFN‐γ (100 U/ml) did not modify their FcγR‐ and CR‐mediated phagocytosis, but significantly up‐regulated FcγRI and CR3 membrane expression as well as their oxidative metabolism and antibody‐dependent cellular cytotoxicity (ADCC). When IL‐10 and IFN‐γ were added simultaneously to neutrophil culture, IL‐10 dose‐dependently reduced IFN‐γ‐induced increase of CR3 expression, O2− production (in response to both FMLP and phorbol 12‐myristate 13‐acetate, or PMA) and ADCC, but did not change FcγRI expression on phagocytes. These results demonstrate that IL‐10 is a significant neutrophil deactivator and provide new information on the role of IL‐10 in the regulation of neutrophil‐mediated inflammatory processes.
Indirect Effects of Oral Tolerance Cannot be Ascribed to Bystander Suppression
Authors: CARVALHO C. R. , VERDOLIN B. A. , VAZ N. M.
Published: November 2003

The addition of tolerated antigens to immunizing doses of unrelated antigens blocks antibody responses to these unrelated antigens. This inhibition, which the authors have called the indirect effects of tolerated antigens, occurs even when the mixture of proteins is injected as soon as 24 h after the oral tolerance induction. The indirect effects also do not require the simultaneous injection of the two proteins: they are still present 72 h after an injection of Ova in Ova tolerant mice, but do not occur if the unrelated protein is injected 24 h before the tolerated protein. In addition, indirect effects do not block secondary responses to unrelated proteins if the primary immunization is made in the absence of the tolerated protein. These results cannot be explained by innocent bystander suppression, which is believed to result from the action of suppressive cytokines released by specific tolerant lymphocytes upon unrelated lymphocytes that would otherwise respond to the second, non‐tolerated antigen. Indirect effects may be better understood in terms of network models.
Influence of Quillaja saponaria Triterpenoid Content on the Immunomodulatory Capacity of Epstein–Barr Virus Iscoms
Authors: DOTSIKA E. , KARAGOUNI E. , SUNDQUIST B. , MOREIN B. , MORGAN A. , VILLACRES‐ERIKSSON M.
Published: November 2003

The immune responses to immunostimulating complexes (iscoms) containing recombinant Epstein–Barr virus (EBV) gp340 envelope protein was evaluated in BALB/c (H‐2d) and CBA (H‐2k) mice. Gp340‐iscoms were used either with a low content of Quillaja triterpenoid adjuvant (L‐iscoms) or supplemented with additional Quillaja adjuvant in the form of iscomatrix (S‐iscoms). Class and subclass distribution of anti‐gp340 antibodies, EBV‐neutralizing antibodies, antigen‐specific T cell proliferation and cytokine production were determined and these results compared to those obtained by immunization with non‐adjuvated gp340. The H‐2d and H‐2k mice were characterized as low or high responders in respect to the level of specific anti‐gp340 antibodies, secretion of IgG2a isotype, antigen‐specific lymphoproliferative capacity, interferon‐γ (IFN‐γ) and interleukin‐10 (IL‐10) production in the basic immunizations with gp340. While presentation of the antigen in iscom formulations with low levels of Quillaja triterpenoids induces a moderate enhancement of the immune responses in the low responder H‐2d mice, supplementation with high levels of iscomatrix immunomodulator was required to enhance the immune responses in the high responder H‐2k mice. In both mouse strains subcutaneous immunization with S‐iscoms resulted in a significant increase of IgG1‐ and IgG2a‐specific antibodies, as well as in strong antigen‐specific proliferative response confirmed by the simultaneous cytokine production. The enhanced antigen‐specific secretion of IL‐2 and IFN‐γ together with the abrogation of IL‐10 and the absence of IL‐4 indicates that the responses were driven towards a Th1‐type rather than Th2‐type immune response. The S‐iscom formulations minimized the differences in immune responses between the two mouse strains, but the capacity of immune sera to neutralize EBV transformation in vitro remained completely strain‐dependent. These data indicate that immune responses generated by iscoms can be manipulated by altering the triterpenoid composition of the iscoms and that the levels of triterpenoids can determine whether or not a Th1‐type response is made.
TNF‐α Regulates GM‐CSF‐, IL‐3‐ or M‐CSF‐Induced Fc ε RII/CD23 Gene Expression and Soluble Fc ε RII Release by Human Monocytes
Authors: HASHIMOTO S. , GON Y. , HAYASHI S. , TOMITA Y. , YODOI J. , HORIE T.
Published: November 2003

The authors examined the regulatory effects of tumour necrosis factor‐α (TNF‐α) on granulocyte macrophage colony stimulating factor (GM‐CSF)‐, interleukin‐3 (IL‐3)‐ or macrophage colony stimulating factor (M‐CSF)‐induced gene expression of the low affinity receptor for IgE (Fc ε RII) on human monocytes and GM‐CSF‐, IL‐3‐ or M‐CSF‐induced soluble Fc ε RII (sFc ε RII) release from monocytes. The expression of GM‐CSF‐, IL‐3‐ or M‐CSF‐induced Fc ε RII on the surface of monocytes was reduced by TNF‐α. The present analysis was designed to examine whether or not TNF‐α could suppress GM‐CSF‐, IL‐3‐ or M‐CSF‐induced Fc ε RII messenger RNA (mRNA) expression and enhance the release of sFc ε RII induced by these cytokines. The addition of TNF‐α to monocyte cultures with GM‐CSF, IL‐3 or M‐CSF significantly reduced Fc ε RII expression on the surface of monocytes and significantly increased sFc ε RII release from monocytes. These results suggest that TNF‐α‐dependent reduction of GM‐CSF‐, IL‐3‐ or M‐CSF‐induced Fc ε RII expression on the surface of monocytes resulted, at least in part, from the suppression of Fc ε RII mRNA and the enhancement of sFc ε RII release.
Polyreactive Binding of Antibodies Generated by Polyclonal B Cell Activation. II. Crossreactive and Monospecific Antibodies can be Generated from an Identical Ig Rearrangement by Differential Glycosylation
Authors: FERNÁNDEZ C. , ALARCÓN‐RIQUELME M. E. , SVERREMARK E.
Published: November 2003

In this work the authors tested the hypothesis that differential glycosylation may be one of the mechanisms by which B cells could be able to produce monospecific or polyreactive antibodies flexibly. The same genetic information will be used in both situations. To prove this hypothesis the authors used mice transgenic for the SP6 (μ+κ) hybridoma cell line producing monoclonal antibodies (MoAbs) with specificity for the hapten 2,4,6‐trinitrophenyl (TNP). In these animals most cells in the periphery express exclusively the transgene SP6. To obtain polyreactive antibodies (Abs) spleen cells from transgenic animals were stimulated in vitro with lipopolysaccharide (LPS). The authors used LPS derived B cell blasts to produce a hybridoma cell collection. A high proportion of the monoclonal antibodies were found to bind to TNP and to crossreact with unrelated antigens. Endogenous immunoglobulins (lgs) were not responsible for crossreactivity since the crossreactive clones only expressed the transgene products. This was demonstrated by polymerase chain reaction (PCR) amplification of cDNA and by sequencing analysis of the PCR products. The nucleotide sequences of the expressed mono‐ and crossreactive genes were identical to the sequences of the rearranged VH and VK SP6 which undoubtedly demonstrates that crossreactive Igs are derived from the same rearrangement and also that no mutations in the VH or VK or in the CDR3 could account for the observed crossreactivity. It is also shown here that the crossreactive antibodies bear the idiotype Id 20‐5 characteristic of SP6 binding Abs. Crossreactive monoclonal antibodies were found to be slightly more glycosylated than the TNP‐monospecific Abs. Furthermore, binding to TNP was not influenced by treatment with tunicamycin, an inhibitor of glycosylation, while, in the same molecule, other types of binding were considerably reduced. This supports the hypothesis of the importance of glycosylation in polyreactivity.
Characterization of Components of the Murine B Cell Receptor Complex and Their Role in Anti‐Ig Stimulation
Authors: VOGEL L. , GO¨TZ M. , HAUSTEIN D.
Published: November 2003

Signal transduction in B cells is mediated by B cell receptor (BCR) complexes that are composed of membrane immunoglobulins (mIg) and additional proteins (e.g. Ig‐α and Ig‐β) that have been implicated with several aspects of B cell activation. In this paper, experiments are described that have been designed to characterize new components of the BCR and to elucidate their involvement in B cell activation directly after the binding of anti‐Ig. The data obtained prove the assumption that IgM is degraded after internalization. Neither immunoglobulin nor the Ig‐α/β heterodimer are re‐expressed after they have disappeared from the cell surface. The newly detected Ig‐associated proteins are neither degradation products of immunoglobulins nor different forms of Ig‐α or Ig‐β. In addition, they are not recognized by antibodies against any of the kinases known to date. The mIg molecules are linked to the cytoskeleton and internalized. On the other hand, the additional Ig‐associated proteins as well as the Ig‐α/β heterodimer could not be detected in the detergent‐insoluble fraction. The former molecules remain on the B cell surface where they are attached to the unaffected isotype and might interact with downstream members of the signalling cascade. In addition, evidence is presented that the internalization of the receptor complex is not necessary for signal transduction and B cell activation. From these results the authors conclude that the BCR complexes are removed from the cell surface after binding of ligands, probably to prevent further activation of this cell, while the Ig‐αssociated proteins interact with the intracellular components of the signal transduction pathway to promote the further activation of the B cells.
Major Histocompatibility Complex Class II Region Confers Susceptibility to Staphylococcus aureus Arthritis
Authors: ABDELNOUR A. , ZHAO YI‐XUE , HOLMDAHL R. , TARKOWSKI A.
Published: November 2003

The importance of the MHC class II region for the development of septic arthritis was studied in a murine model of haematogenously induced Staphylococcus aureus arthritis. In the first experiment MHC class II deficient mice (Aβ−/−) and their heterozygous (Aβ+/−) littermates were intravenously inoculated with a single dose of toxic shock syndrome toxin‐1 producing S. aureus LS‐1 strain. The results demonstrate that the expression of class II MHC molecules increases the prevalence and severity of arthritis. To analyse the impact of MHC class II haplotypes on the disease onset and progression the authors used congenic C3H.NB, C3H.Q and C3H/HeJ mice in the second set of experiments. The results show that C3H/HeJ mice developed the highest frequency and the most severe course of arthritis compared with C3H.NB and C3H.Q animals. Immunohistochemical analysis of arthritic joints revealed equal number of macrophages, CD4+ and CD8+ lymphocytes in the inflamed synovia in all the congenic mice. In contrast, the number of MHC class II expressing cells was higher in the arthritic joints of C3H/HeJ mice compared with the congenic strains (P<0.001). Furthermore, serum levels of proarthrtitogenic cytokines, such as tumour necrosis factor and interleukin‐6 were higher in C3H/HeJ group. This study indicates that MHC class II expression is necessary for the development of S. aureus arthritis in mice and that different MHC class II haplotypes confer varying susceptibility for development of joint inflammation induced by staphylococci.
Diminished Contact Hypersensitivity Response in IL‐4 Deficient Mice at a Late Phase of the Elicitation Reaction
Authors: WEIGMANN B. , SCHWING J. , HUBER H. , ROSS R. , MOSSMANN H. , KNOP J. , RESKE‐KUNZ A. B.
Published: November 2003

Contact hypersensitivity (CHS) is thought to depend on the activation of T cells of Th1 and/or Tc1 type. The role of Th2/Tc2 cells in the contact allergic reaction is not clear. The aim of this study was to analyse the functional contribution of Th2/Tc2 cells in CHS using the interleukin‐4 (IL‐4) deficient mouse model. Interleukin‐4 deficient (IL4T) and control (wt) mice were sensitized by epicutaneous application of 2,4‐dinitrofluorobenzene. The ear swelling response measured 24 h after challenge was similar in IL4T and control mice. However, from 48 h onwards, ear swelling values were significantly reduced in IL4T mice. The stimulatory capacity of freshly isolated as well as 3‐day cultured epidermal cells, prepared from IL4T and wt mice, for allogeneic T cells in a primary and secondary response, was comparable. The reduced number of T cell receptor (TCR) γδ+ cells observed in epidermal sheets prepared from IL4T mice was not responsible for the decreased ear swelling response in IL4T mice, because the use of TCR δ deficient mice lacking TCR γδ+ cells revealed a down‐regulatory role of this cell population in the CHS response. The data indicate that the effector stage of the CHS response can be subdivided into two phases. The first phase proceeds efficiently in IL‐4 deficient mice indicating the dependence on Th1/Tc1 cells, while the second phase does not develop in mice lacking IL‐4, suggesting the possibility that Th2/Tc2 cells intensify the reaction.
Progressive Accumulation of Bacterial Lipopolysaccharide In Vivo During Murine Acute Graft‐Versus‐Host Disease
Authors: PRICE K. S. , NESTEL F. P. , LAPP W. S.
Published: November 2003

In a previous report the authors demonstrated that acute graft‐versus‐host disease (GVHD) was associated with pathologic amounts of tumour necrosis factor alpha (TNF‐α) and the appearance of lipopolysaccharide (LPS) in the blood of GVH reactive mice just prior to death. In this study the authors have investigated the kinetics of LPS accumulation in different organs during the course of acute GVHD using a murine model. Unirradiated C57BL/6×AF1 (B6AF1) mice were transplanted with C57BL/6 (B6) lymphoid cells and killed at predetermined times after transplantation for LPS analysis. Control animals were injected with either 60×106 B6AF1 lymphoid cells (syngeneic) or 60×106 irradiated (2000 rad) CBA lymphoid cells (allogeneic). Lipopolysaccharide began to appear in the liver and the spleen of GVH reactive mice from day 2 post‐transplant and by day 10 all GVH reactive mice tested positive for hepatic and splenic LPS. Low levels of LPS were detected in some control mice from days 2 to 10 post‐transplant but LPS was never detected after day 10 in control groups. Total hepatic and splenic LPS in acute GVH reactive mice peaked at a time coincident with the appearance of LPS in the serum and with the onset of mortality. These results demonstrate that tissue levels of LPS increase throughout the course of acute GVHD and are sufficient to trigger the release of pathologic amounts of TNF‐α from primed macrophages resulting in the cachexia and mortality associated with acute GVHD in this model.
Serological Diagnosis of Trypanosoma rangeli Infected Patients. A Comparison of Different Methods and its Implications for the Diagnosis of Chagas' Disease
Authors: VÁSQUEZ J. E. , KRUSNELL J. , O¨RN A. , SOUSA O. E. , HARRIS R. A.
Published: November 2003

Venous blood from 65 Panamanian schoolchildren living in an area endemic for both Trypanosoma cruzi and T. rangeli were screened for the presence of these parasites. Trypanosoma rangeli were isolated and cultured from four individuals. Serological tests of all 65 sera were performed, including immunohaemagglutination (IHA), indirect immunofluorescence assay (IF) and ELISA using both T. rangeli and T. cruzi as antigens, as well as T. cruzi synthetic peptides in an ELISA assay. Results indicated a higher immunoreactivity to T. rangeli preparations than to T. cruzi within the studied population, which could be divided into four ‘serological responder’ groups. Interestingly, the panel of SAPA and other T. cruzi synthetic peptides were not useful in the discrimination of patients. Furthermore, patients from whom parasites had been isolated could not be distinguished from those of two other groups. Significant immunoreactivityto T. cruzi preparations was displayed in all responder sera, despite total lack of evidence of infection with this parasite. The immunobiological significance of T. rangeli infection is unclear, but these data indicate that it is a compounding problem in the accurate diagnosis of pathological T. cruzi infection by serological analysis. The relationship of these cohabiting species, in respect to infection outcome and immunological activation, is discussed.
Altered IL‐1β and IL‐2 mRNA Expression in Murine Splenic Cells After Concomitant Stimulation by Semliki Forest Virus and Lipopolysaccharide
Authors: YANG J. , HUKKANEN V. , SELJELID R. , SALMI A. A.
Published: November 2003

Co‐infection with virus and bacteria happens frequently and often results in an exacerbated clinical course of the disease, possibly due to mechanisms including altered cytokine production. In the present study, the authors investigated the combined effects of avirulent Semliki Forest virus (SFV‐A7) and bacterial lipopolysaccharide (LPS) on the interleukin‐1β (IL‐1β) and IL‐2 gene expression in murine splenic cells. The authors found that 10 ng/ml of LPS in the culture medium induced expression of IL‐1β but not IL‐2, while infection with SFV‐A7 did not induce either of these two cytokines. However, when SFV‐A7 and LPS were applied together, a synergistic increase of both IL‐1β and IL‐2 was observed. Further experiments showed that addition of SFV‐A7 3 h before LPS enhanced, whereas addition of the virus 3 h after the LPS inhibited, IL‐1β gene expression. These results indicate that an interaction of virus and Gram‐negative bacteria can result in an altered cytokine gene expression.
Mycophenolate Mofetil, Azathioprine and Cyclophosphamide Enhanced Efficacy Combined with Cyclosporine in Rat Cardiac Transplantation
Authors: O¨straat O¨ , Qi Z. , Olausson M. , Tufveson G. , Ekberg H.
Published: November 2003

Anti‐proliferative drugs have been used for immunosuppression since the introduction of clinical transplantation. Most transplant centres include azathioprine (Aza) and cyclosporine (CyA) in their standard regimens, despite several controlled studies having failed to confirm the benefit of this combination. Aza is still the most commonly used anti‐proliferative drug, although no major differences in immunosuppressive or toxic effects have been shown between Aza and cyclophosphamide (Cph). Cph as an adjunct to CyA has never been tested in a randomized study. Recently, mycophenolate mofetil (MMF) has been developed as the most selective inhibitor of T‐ and B‐cell proliferation and promoted as an adjunct to CyA treatment. In the present study, the additive or synergistic effects of these three anti‐proliferative agents in combined treatment with CyA have been investigated using a rat cardiac transplantation model in which the immunomodulator linomide (Lin) was included as a potentiator of rejection. As single drug treatment, CyA, Cph and MMF, but not Aza, exerted a beneficial effect on graft survival. This prolongation of graft survival was abrogated when any one drug was administered together with Lin. The addition of MMF, Aza or Cph to CyA plus Lin treatment improved the graft survival significantly, thus demonstrating each of the anti‐proliferative drugs to exert additive or synergistic effects in conjunction with cyclosporine. MMF seemed to be the most effective and least toxic of the drugs tested.
Degradation of Amyloid A Precursor Protein SAA by Macrophage Cell Lines Obtained from Amyloid Resistant and Susceptible Strains of Mice
Authors: HAM D. , CAOURAS V. , RADZIOCH D. , GERVAIS F.
Published: November 2003

Reactive AA amyloidosis can be induced in mice in a model of sustained inflammation following daily casein subcutaneous injections. However, the development of AA amyloidosis is known to vary in different strains of mice. The C57BL/6 strain is susceptible to the development of amyloidosis while the A/J strain is resistant. The degradation of purified serum amyloid A (SAA) protein by human monocytes as well as by mouse macrophages has been shown. The resistance/susceptibility of different mouse strains to the development of systemic amyloidosis may therefore be related to a difference in the ability of macrophages to degrade SAA. The authors have used bone marrow‐derived macrophage cell lines obtained from susceptible C57BL/6 (ANA‐1) and resistant A/J (A/J 10) mouse strains to compare their ability to degrade HDL‐SAA in vitro. Cells were incubated with HDL‐SAA for up to 72 h and the culture medium was analysed by SDS‐PAGE to determine the rate of SAA degradation by the macrophages. The A/J 10 cells (resistant) were found to initiate a constant HDL‐SAA degradation promptly whereas ANA‐1 cells (susceptible) showed an intermittent block in the degradation of the precursor. Activation of macrophages by lipopolysaccharide (LPS) or interferon‐γ (IFN‐γ) hampered the precursor degradation suggesting that the activation process may favour extracellular accumulation of the precursor leading to a partial degradation and fibril formation.