bluuebunny/crossref_metadata_2025 huggingface dataset:
Ocular and Systemic Determination of and Tumour Necrosis Factor in a Patient with Ocular Inflammation
Authors: PALEXAS G. N. , SUSSMAN G. , WELSH N. H.
Published: July 2006

Cytokines such as interleukin‐1β (IL‐1β) and tumour necrosis factor (TNF) may play an important role in ocular inflammation. We studied a patient with clinical features of sympathetic ophthalmia secondary to previous penetrating ocular injuries, and compared the ocular and systemic levels of IL‐1β and TNF to control serum, and correlated these findings to histopathological sections of the patient's eye. Histology showed the present‐c of a diffuse chronic inflammatory infiltrate within the choroid and in a perivascular distribution in the retina. The significantly elevated ocular and systemic levels of IL‐1β and TNF suggest that there is not only a localized ocular response hut a systemic response as well. The presence of IL‐1β TNF may play a role in the pathogenesis of ocular inflammation once the blood ocular barrier has been breached and ocular antigens have been exposed to the systemic immune system.
Enhanced Spontaneous and Induced LAK Function in Baboons Receiving Total Lymphoid Irradiation (TLI)
Authors: GRAY C. M. , STARK J. H. , SMIT J. A. , MYBURGH J. A.
Published: July 2006

TLI has potent immunosuppressive effects, but there is concern over its possible carcinogenic properties. The aim of the study was to assess to what degree TLI may compromise natural killer (NK) and lymphokine activated killer (LAK) cell function. There was a significant increase (p < 0.0001) in both NK‐cell function (measured against K562 targets) and spontaneous LAK‐cell function (measured against DAUDI targets) in fresh blood lymphocytes throughout a course of 8 × 100 cGy fractionated TLI. This may be related to a three‐ and sevenfold increase, respectively, in CD16+ CD8‐ and CD56+ CD2‐ cell frequencies over the same period. Mitogen‐induced interleukin 2 (IL‐2) synthesis from blood lymphocytes was inhibited by up to 75% with as little as 100 cGy of TLI. Expression of IL‐2 receptors on fresh lymphocytes did not vary and remained low. Therefore spontaneous LAK occurrence appeared to be triggered through an lL‐2‐independent pathway. The in vitro addition of IL‐2 verified that cells retained their ability to respond to this lymphokine resulting in greatly enhanced induced LAK function. This was most probably mediated by CD56+ cells which were found to readily express IL‐2 receptors upon mitogen stimulation. In conclusion, fractionated low‐dose TLI appears to enhance MHC unrestricted immune surveillance in a manner independent of IL‐2 production.
Generation of Lymphokine‐Activated Killer Cells by Adherent LGL Phenotype Cells and Non‐Adherent T Lymphocytes
Authors: OUESLATI R. , MAALEJ M. , CHOUIKHA M.
Published: July 2006

In this work we separated human blood lymphocytes (PBL) in two populations (A‐LAK and NALAK cells) by the adherence plastic method. A maximum adherence of cells was obtained after 2 days of PBL incubation in LAK medium containing 500 U/ml rIL‐2. The A‐LAK cells had LGL phenotype but 40% of them had a macrophage phenotype marker and less than 20% weakly expressed a T‐cell marker. This population, when reincubated in culture, produced an increasing titre of interferon. At the same time, a significant NK activity against K562 target cells was measured just after enrichment; these enriched adherent cells also developed an increased LAK activity against DAUDI cell lines, ninefold more at 6 days than when assayed just after enrichment. In contrast, 75% of the NA‐LAK enriched cells expressed T‐cell marker; these produced two‐ to threefold less interferon than A‐LAK cells at all time‐points. The NA‐LAK lymphocytes enhanced principally LAK activity measured by 70% lysis against DAUDI target cells tested at 6 days of culture. Further studies are in progress to determine the nature of the effector cells that mediated LAK activity.
Immunological Consequences from Exposure to Benzo(a)pyrene During Pregnancy
Authors: URSO P. , ZHANG W. , COBB J. R.
Published: July 2006

Progeny and maternal immune status after benzo(a)pyrene (BP) exposure of mothers at midpregnancy is disrupted in fetal liver (FL), in spleen and in thymus during pregnancy and postnatally. Mice suffer deficiencies in splenic and thymic mixed lymphocyte responses (MLR), and disorientations of T antigen expressing cells, punctuated by exorbitant increases in Lyt2, especially in FL, FL Lyt2 do not suppress an MLR, while Lytl mediate suppression. Isolated Thyl show a weak response to Concanavalin A: FL Thyl weakly express an MLR. Maternal macrophages and progeny B cells are also functionally abnormal. Thus, BP Induces generalized immune deficiency that may affect ontogeny and which is potentially deleterious to health.
Iron Deficiency and Immune Responses
Authors: LATUNDE‐DADA G. O. , YOUNG S. P.
Published: July 2006

Iron deficiency is often associated with the impairment of the immune system, particularly the functioning of T‐cell lymphocytes. Studies were therefore carried out in mice to investigate the involvement of iron in T‐cell functions. Iron deficiency in mice was induced nutritionally by feeding diets differing in iron content for 4 weeks to produce normal, moderately low and severely low haematological indices. The proliferation of lymphocytes stimulated by Con A from these groups of mice was significantly affected by the degree of iron deficiency. Iron depletion also resulted in decreased production of interleukin 2 (IL‐2) in the proliferating cells. The implications of these findings in the manifestation of iron deficiency anaemia are discussed.
IgG Subclass Values from Normal Children in Cape Town
Authors: GODDARD E. A. , MALAN E. H. , BEATTY D. W.
Published: July 2006

Sensitive and reproducible enzyme‐linked immunoabsorbent assays (ELISA) have been developed to quantitate IgG subclass levels using monoclonal antibodies. Normal values for serum IgG subclass levels were determined in 300 healthy children between 6 months and 14 years of age and in SO adults. High levels of IgG 1 and delayed maturational development of IgG 2 in children from Cape Town are different to results reported from developed countries. Genetic differences may account for this.
Anergy and Suppression in B‐Cell Responses
Authors: ELLIOTT J. I.
Published: July 2006

Two main ideas have been put forward to explain the unexpectedly low anti‐hapten antibody titres which can result from pre‐priming a mouse with carrier before hapten‐carrier immunization. The first involves the interaction of a network of idiotype‐specific suppressor T cells, the second instead arguing for the role of intrinsic B‐cell anergy. This paper proposes that the data available can equally be interpreted as reflecting the suboptimal interaction between T and B cells at differing stages of maturity, provided that memory B cells can be divided into two subsets. Further, it is suggested that these considerations must be taken into account in the analysis of B‐cell anergy in receptor transgenic mice.
Primary Stimulation of CD4+ Cells in the Presence of IL‐4 or IFN‐γ Alters the Frequencies of Cytokine‐Producing Cells at Restimulation
Authors: CARDELL S. , SANDER B. , MÖLLER G.
Published: July 2006

The induction of specific effector functions in naive T cells may be directed by accessory signals during activation. These could be elicited through binding to cell surface molecules or through factors secreted by antigen‐presenting cells or other simultaneously activated cells. We have investigated the influence of CD8+ cells and of exogenousiy added cytokines (interleukin (IL)‐2, IL‐4 and interferon (IFN)‐γ) on the cylokine production in splenic CD4+ T cells. IL‐2, IL‐4, IL‐5 and IFN‐γ production in CD4+ cells was measured at the single cell level during primary mitogen stimulation in vitro in the presence or absence of factors or CD8+ cells. On day 5 the cells were restimulated with mitogen alone and analysed to evaluate the short‐term development of cytokine‐producing cells in such cultures. Preactivation in the presence of either exogenous IL‐4 or IFN‐γ led to an increased production of IL‐4 and IFN‐γ respectively at restimtilation, and the effects of both IL‐4 and IFN‐γ were augmented by IL‐2. After preactivation in the presence of IL‐2 and IL‐4, every third CD4+ cell could be induced to produce IL‐4. Exogenous IL‐4 or IFN‐γ further decreased each other's production. Depletion of CD8+ cells before activation resulted in a slight increase of IL‐4‐producing cells, indicating that simultaneous activation of CD8+ cells will influence lymphokine production in CD4+ cells. The results suggest that the pattern of lymphokines induced in naive cells may be influenced by factors secreted by preactivated CD4+ and CD8+ cells, and that naive cells are preferentially ‘recruited’ to produce similar cytokines.
Anti‐Golgi Antibody in Rheumatoid Arthritis Patients Recognizes a Novel Antigen of 79 kDa (Doublet) by Western Blot
Authors: HONG H. S. , MORSHED S. A. , TANAKA S. , FUJIWARA T. , IKEHARA Y. , NISHIOKA M.
Published: July 2006

We have detected cytoplasmic anti‐Golgi antibody (AGA) during a routine immunofluorescence test for detecting autoantibodies. Two sera from patients with rheumatoid arthritis (RA) reacted to the Golgi complex by an indirect immunofluorescence technique on HEp‐2 cells. Localization of AGA in the Golgi complex was confirmed by double‐staining with antibodies to β‐COP. The effect of monensin on the integrity and morphology of the Golgi complex was also studied. To confirm the presence of AGA further, we performed immuno‐electron microscopy. Both sera reacted with cytoplasmic antigen located in the Golgi complex of various animal tissues. Furthermore, by using the Western blot technique, both sera reacted to a relative molecular weight (MW) of 79 kDa (doublet) Golgi antigen purified from rat liver. To our knowledge, this study may be the first to identify the relative MW of Golgi antigen by the Western blot method. Identification of this antibody could provide better understanding of protein synthesis and secretion. The presence of AGA in RA patients further substantiates the diversified nature of autoantibody production seen in this disease.
Adhesion of Subsets of Human Blood Mononuclear Cells to Endothelial Cells In Vitro, as Quantified by Flow Cytometry
Authors: BENSCHOP R. J. , DE SMET M. B. M. , BLOEM A. C. , BALLIEUX R. E.
Published: July 2006

Binding of leucocytes to endothelial cells (EC) is essential as an initial step in inflammatory responses. We present a rapid, non‐radioactive method to measure adhesion of human lymphoid cells to EC using flow cytometry. Freshly isolated peripheral blood mononuclear cells (PBMC) were allowed to adhere to EC grown in 24‐well plates. Non‐adhering cells were removed, after which adhering cells and EC were dissociated using trypsin/EDTA. These samples were subsequently analysed by flow cytometry, using scatter properties to distinguish between adhering cells and EC. The ratio of the number of adhering leucocytes and EC was calculated to quantify adhesion. Results of the flow cytometric adhesion assay were comparable to those obtained with a conventional adhesion assay using chromium‐labelled cells. We additionally show that by using the flow cytometric adhesion assay, adhesion of lymphocyites and monocytes present within the adhering PBMC can be quantified simultaneously. As a model, the contribution of LFA‐1 (CD11a,CD18) and ICAM‐1 (CD54) in adhesion of PBMC to EC was studied. It was found that adhesion of lymphocytes and monocytes is regulated differently by phorbol ester and that the relative contribution of LFA‐1 and ICAM‐1 differs for both cell types.
Studies of Antibody to Herpes Simplex Virus Fcγ‐Binding Protein gE in Patients with Rheumatoid Arthritis, Juvenile Rheumatoid Arthritis and Normal Controls
Authors: WILLIAMS R. C. , MALONE C. C.
Published: July 2006

IgG antibody to gE, the Fcγ‐binding herpes simplex 1 (HSV‐1) viral glycoprotein, was studied in 49 rheumatoid arthritis (RA) patients and 43 normal controls. Antibody to gD, another important HSV‐1 antigen, was assayed in parallel. No difference between RA patients and normal controls was found in levels of anti‐gE antibody measured by reactivity of IgG F(ab')2 fragments reacting with gE coated to ELISA plates. No difference in anti‐gD antibody was recorded between normals and patients with RA. Levels of IgG anti‐IgE antibody did not correlate with quantitative elevations of serum rheumatoid factor (RF) in RA patients. When IgG anti‐gE and anti‐gD were assayed in 20 patients with juvenile rheumatoid arthritis and 22 children controls, no significant differences were noted. However, when individual RFs from patients with RA were tested for reactivity against a panel of affinity‐isolated E(ab')2 antibodies to gE, some evidence for individual autospecificity was obtained. Four of 20 monoclonal IgM RFs produced from RA patients' B cells showed marked elevations of reactivity with some RA patients'F(ab')2 antibodies to gE. All four of the monoclonal RFs showing this specificity were derived from RA synovial tissue B cells. These findings may provide support for the concept that some RFs in patients with RA show individual specificity for internal image determinants of IgG antibodies to viral Fcγ‐binding proteins.
Induction of Cellular Immune Reactions by A36, an Antigen Complex of Mycobacterium paratuberculosis: Compariso of A36 and Johnin Components
Authors: GILOT P. , DE KESEL M. , COENE M. , COCITO C.
Published: July 2006

Paratuberculosis (Johne's disease) is a chronic enteritis syndrome of ruminants, which is due to infection by Mycobacterium paratuberculosis. Cutaneous testing with proteins extracted from a mycobacterial culture fluid (johnin‐PPD) is currently used to evaluate the cellular immune status. We have compared the components of johnin‐PPD with those of the A36 complex, a thermostable macromolecular antigen (TMA) present in the cytoplasm and associated with the cell wall of M. paratuberculosis. The presence in the johnin‐PPD of fifteen A36 components has been shown by Western blotting. Moreover, monoclonal antibodies, which bind respectively to the 65‐kDa M. leprae heat shock protein, the 28‐kDa M. leprae superoxide dismutase, and M. tuberculosis lipoarabinomannan, recognized components of the johnin‐PPD. The ability of A36 to trigger delayed hypersensitivity reactions in sensitized rabbits, and to induce the proliferation of T lymphocytes from the lymph nodes of A36‐sensitized mice, matched that of johnin‐PPD. The homology levels of T epitopes between A36 and the TMA complexes of M. phlei, M. bovis, M. tuberculosis and M. avium were estimated, in a lymphoproliferation assay, to be 51, 52, 59 and 94% respectively. A strong cross‐reactivity of A36 with an M. leprae sonicate was also observed by cutaneous testing. The A36 components within the 45.2–26.8‐kDa and the 21.6–19.8‐kDa ranges were proved to induce the proliferation of T lymphocytes from sensitized mice. This work supports the possible use of the A36 complex, and of some of its components, for cutaneous tests and lymphocyte proliferation assays, in order to monitor cellular immunity in Johne's disease.
Restricted Expression of Immunoglobulin Light Chain mRNA and of the Adhesion Molecule CD11b on Circulating Monoclonal B Lineage Cells in Peripheral Blood of Myeloma Patients
Authors: JENSEN G. S. , BELCH A. R. , KHERANI F. , MANT M. J. , RUETHER B. A. , PILARSKI L. M.
Published: July 2006

Circulating monoclonal B cells in peripheral blood from patients with multiple myeloma or with monoclonal gammopathy of undetertmined siginificance (MGUS) have previously been shown to express CD19, CD20, and PCA‐1 and are predominantly CD45R0+, characterizing them as very late stage B cells. This work shows that the abnormal B cells are monoclonal as defined by their exclusive expression of either K or λ light chain mRNA, and that the same type of light chain mRNA is expressed in both bone marrow plasma cells and blood B cells. These abnormal tumour related circulating B cells express high densities of CD11b, a β2‐integrin, which is expressed in a conformationally active state as defined by reactivity with monoclonal antibody 7E3. Normal peripheral blood B cells which do not bear CD11b acquire a high density after stimutation with pokeweed mitogen (PWM). At day 4 of culture, the expression of CD11b on normal CD19+ B cells was nearly comparable to that of the circulating myeloma late stage B cells. After PWM stimulation of circulating myeloma B cells the expression of CD11b was gradually lost during 4 days of culture, suggesting that its expression is dynamically regulated. Two patients with no phenotypically abnormal B cells in their blood at diagnosis acquired a large subset of CD11b+ B cells 4 weeks after initiation of chemotherapy. In most patients, a subset of the circulating myeloma B cells express a low density of CD5. The proportion of CD19+ B cells in the bone marrow expressing CD11b was much reduced compared with peripheral blood B cells, and CD11b was not detectable on plasma cells in the bone marrow, suggesting a sequential relationship of the B‐cell subsets detected in our population of patients, involving gradual loss of CD11b concurrent with the loss of CD19 during B lineage differentiation. These cells appear to represent a continuously differentiating monoclonal B lineage culminating in the CD11b plasma cell entrenched in the bone marrow. We speculate that the expression of conformationally active CD11b on the abnormal B cells in peripheral blood mononuclear cells of myeloma patients facilitates transendothelial migration of circulaling myeloma B cells to the bone marrow.
Spontaneous In Vivo Gene Transcription of Interleukin‐2, Interleukin‐3, Interleukin‐4, Interleukin‐6, Interferon‐Gamma, Interleukin‐2 Receptor (CD25) and Proto‐Oncogene c‐myc by Rheumatoid Synovial T Lymphocytes
Authors: WAALEN K. , SIOUD M. , NATVIG J. B. , FØRRE Ø.
Published: July 2006

Rheumatoid synovial T lymphocytes were investigaled for the presence of mRNA for the cytokines interleukin‐2, ‐3, ‐4, ‐6, interferon‐gamma, the interleukin‐2 receptor (CD25) and the proto‐oncogene c‐myc. The isolated RNAs were analysed by dot blot and Northern blot hybridization. Our results show that synovial T lymphocytes from patients with rheumatoid arthritis (n= 12) had spontaneous tn vivo gene transcription of interleukin‐2 (93%), interleukin‐4 (67%), interleukin‐6 (92%), interleukin‐2 receptor (92%) and the proto‐oncogene c‐myc (67%). Only a few of the RA patients had synovial T cells with increased expression of mRNA for interleukin‐3 (25%) and interferon‐gamma (25%). The amounts of mRNA for the various cytokines and activation molecules produced by the rheumatoid synovial T lymphocytes were in most instances comparable to those of normal peripheral blood T lymphocytes activated in vitro by the mitogen phytohaemagglutinin. The data thus indicate that the synovial T lymphocytes are activated in viov in the majority of rheumatoid arthritis patients.
A Sensitive Simple ELISA for Quantitation of Sensitizing IgG from Dissolved Erythrocytes
Authors: GYIMESI E. , KÁVAI M. , CSÍPÖ I. , SZEGEDI Gy.
Published: July 2006

A simple enzyme‐linked immunosorbent assay has been developed for quantitating the rabbit IgG present on the surface of sensitized red blood cells. A suitable cell lysis was carried out by an alkaline buffer. which dissolved the erythrocytes without forming any precipitate and without disruption of IgG, and facilitated the dissociation of the immune complexes, i.e. the erythrocyte anti‐erythrocyte rabbit IgG. In this alkaline buffer of pH 11.4 the IgG adsorbed directly into wells of microtitration plates unprecoated with anti‐rabbit IgG. The detection was performed with alkaline phosphatase‐conjugated goat anti‐rabbit IgG and p‐nitrophenyl phosphale as substrate. Constructing a calibration curve from rabbit IgG made possible the calculation of molecules of IgG bound per red cell. The method was sensitive for the detection of fewer than 500 bound IgG molecules per erythrocyte.
Oligonucleotide Probes to the 5’End of the Framework 3 (FR3) Gene Segment detect Polymorphisms of VH Gene Sequences encoding Biologically Important Amino Acid Residues
Authors: RUBINSTEIN D. B. , SYMANN M. , GUILLAUME T.
Published: July 2006

In this study, we demonstrate the use of oligonucleotide probes to the most VH family‐speeific framework, the Eramework 3 (ER3) gene segment, in the analysis of germline DNA. Compared with the hybridization of restriction enzyme‐digested genomic DNA to large, cloned human VH family probes, hybridization with oligonueleotides derived from nucleotides 199 to 258 of the ER3 gene segments both yields less complex patterns and highlights polymorphic variations between individuals. The potentially important role played by amino acid residues encoded by the FR3 gene segment in influencing conformation, antigen binding and affinity of the physically proximal complementarity determining regions (CDRs) suggests FR3 sequence conservation to be biologically significant. Genomic studies combining information from both CDR and framework restriction fragment polymorphisms can prove useful in detecting specific genes and in identifying haplotypes of interest.
HLA Class II+ Human Keratinocytes present Mycobacterium leprae Antigens to CD4+ Thl‐Like Cells
Authors: MUTIS T. , DE BUEGER M. , BAKKER A. , OTTENHOFF T. H. M.
Published: July 2006

In a variety of inflammatory skin diseases like leprosy, keratinocytes (KC) are induced to express MHC class II molecules and may therefore serve as antigen‐presenting cells (APC) for MHC class II restricted T cells infiltrating the lesions. However, KC have been thought to be improper APC for MHC class II restricted T cells and to drive T cells into an anergic rather than into an activation state. We evaluated this issue in relation to leprosy and tested whether HLA‐DR+ KC could present M. leprae antigens to well‐defined, CD4 +, cytotoxic as well as proliferative, Thl ‐like cell clones. Using a recently developed sensitive assay system which employs intact layers of basal KC as APC we found that most T‐cell clones (6/8) lysed HLA‐DR+ KC pulsed with M. leprae antigens. KC were only recognized after induction of HLA‐DR expression by IFN‐γ, in an antigen‐specific and HLA class II restricted manner. All T‐cell clones tested also showed significant proliferation and IFN‐γ production in response to M. leprae antigens presented by HLA‐DR+ KC, arguing against a KC dependent anergizing effect on T cells. Thus, HLA class II+ KC can function as proper APC for HLA class II restricted CD4+ Th l ‐like cells. It seems therefore possible that antigen presentation by KC contributes to the local cell‐mediated immune responses in DTH lesions.
Mechanisms whereby Lytic Granules from Cytotoxic T Lymphocytes Damage Guinea Pig Ventricular Myocytes
Authors: BINAH O. , KLINE R. , BERKE G. , HOFFMAN B.
Published: July 2006

During immunological rejection of the transplanted heart, cytoloxic T lymphocytes (CTL) infiltrate the myocardium and by damaging the myocytes contribute to loss of function. To address one important aspect of heart transplant rejection, we investigated in guinea pig ventricular myocytes how CTL‐derived lytic granules containing the pore‐forming protein perform reduce the membrane potential (Vm) and cause cell damage. The reduction in VM was biphasic; within 8.4 × 1.9 min VM was reduced from a control value of ‐78.4 × 1.9 mV to ‐69.9 × 3.5 mV. Subsequently, within 6.7 × 2.1 min VM declined to ‐ 3.4 × 1.2 mV. associated with a progressive contracture. Under whole cell voltage clamp, in myocytes held at their resting potential (VM= ‐76.2 × 0.9 mV), granules induced discrete inward current steps (resembling “single channel’activity), with a mean amplitude of ‐ 86.8 × 1.4 pA and open times lasting from seconds up to several minutes. The mean conductance and reversal potential calculated from the linear regression analysis of the I‐V relations were 1390 pS and ‐ 6.8 mV. respectively. The probable non‐selectivity of these “channels” and the resultant loss of membrane K+ selectivity can account for the reduction in VM. At the same time, opening of these pores leads to Ca‐+ overload, resulting in contracture and cell damage.
Membrane‐Incorporated Surrogate Receptors Increase Antigen Presentation by Resting B Cells, but not by LPS‐Activated Blasts
Authors: GONTIJO C. M. , MÖLLER G.
Published: July 2006

Palmitate‐conjugated monoclonal antibodies specific to ovalbumin were itiserted into the cell membrane of normal resting B cells and LPS‐activated blasts. These two decorated B cells were tested for their ability to act as antigen‐presenting cells for ovalbumin‐specific I‐Ad‐restricted T‐cell hybridomas. It was found that the antibody‐decorated resting B cells presented antigen more efficiently than non‐decorated controls. However, no increment was observed when decorated LPS blasts were compared with non‐decorated blasts. This is explained by the fact that the inserted antibodies quickly disappeared from the cell membrane of LPS blasts, while they were retained fora long period in the membrane of resting B cells.
Gastric Interleukin‐8 and IgA IL‐8 Autoantibodies in Helicobacter pylori Infection
Authors: CRABTREE J. E. , PEICHL P. , WYATT J. I. , STACHL U. , LINDLEY I. J. D.
Published: July 2006

Gastric infection with Helicobacter pylori is frequently characterized by neutrophil infiltration. The production of the neutrophil‐activating peptide (NAP‐1/IL‐8) and mucosal IgA autoantibodies to IL‐8 by human antral biopsies have been examined during short‐term in vitro culture. Detectable IL‐8 was secreted by 84% of H. pylori‐negative patients with normal antral mucosa (range <0.07–61.5 ng/mg biopsy protein, n=19). Concentrations in 4 patients with reactive gastritis and 10 with inactive gastritis were not significantly different from subjects with normal mucosa. In H. pylori‐positive patients with active gastritis and neutrophil infiltration into the epithelium (n=17) IL‐8 secretion was significantly increased relative to subjects with normal mucosa (p > 0.0001), inactive gastritis (p <0.001) and reactive gastritis (P<0.01). IL‐8 concentrations in active gastritis were significantly correlated with the extent of epithelial surface degeneration (r=0.64). IgA autoantibodies were present in 19 patients (13 active, 4 inactive gastritis) and concentrations were significantly correlated with IL‐8 production (p<0.001). Gastric synthesis of IL‐8 is likely to be an important factor in regulating mucosal neutrophil infiltration and activation in patients with H. pylori infection. The local production of IgA antibodies to IL‐8 may represent a down‐regulatory response of the host to limit mucosal damage associated with a chronic bacterial infection.
Regulation of Natural Killer Cell Activity by Transforming Growth Factor‐β and Prostaglandin E2
Authors: MALYGIN A. M. , MERI S. , TIMONEN T.
Published: July 2006

The separate and combined effects of transforming growth factor‐β1 (TGF‐β1) and prostaglandin E2 on human natural killer (NK) activity were studied. Peripheral blood lymphocytes (PBL) and large granular lymphocytes (LGL, 70–90% purity) were used as effector cells and K562 as targets. Overnight incubation of the effector cells with TGF‐β1 resulted in a significant inhibition of NK activity. TGF‐β1 did not influence the expression of CD3, CD 16, CD 18 or CD56 antigens on PBL. Combination of TGF‐βl with indomethacin gave the same NK‐suppressive effect as TGE‐β1 alone, showing that the inhibition of NK acuvity by TGF‐β1 is not due to an increase in PGE2 levels. TGF‐β did not influence cAMP level in PBL whereas PGE2 significantly increased it. On the other hand. TGE‐β1 and PGE2 showed an additive inhibitory effect on NK activity. TGF‐β1 did not reduce the binding of PBL and LGL to K562. PGE2 suppressed the binding and TGF‐β1 did not influence this suppression. TGF‐β1 also suppressed IL‐2‐induced activation of NK activity and increase of expression of the granule proteins granzyme A and perforin. PGE2 did not appear to affect granzyme A and perforin contents. The results indicate that TGF‐β1 and PGE2 suppress NK activity by different mechanisms.
Anti‐Tumour Activity of Idiotype‐Specific, MHC‐Restricted Th1 and Th2 Clones In Vitro and In Vivo
Authors: LAURITZSEN G. F. , WEISS S. , BOGEN B.
Published: July 2006

Idiotypes (Id) can serve as individual markers on B cells; therefore, cytotoxic Id‐specific T cells may play a significant role in immunological surveillance of Id+ B‐cell tumours. We have investigated the anti tumour activity of CD4+ BALB/c Thl and Th2 clones which recognize a processed Id of the syngeneic λ2315 L chain in the context of the class II MHC molecule I‐Ed. Id‐specific T cells and A 20/46 B lymphoma cells transfected with the λ2315 gene were injected s.c. into the same site of BALB/c mice (Winn assay). The results show that both Th l and Th2 clones can protect against tumour development. The protection was Id‐specific because T cells did not influence tumour development by an A20/46 B lymphoma cell line transfected with the pSV2neo expression vector alone. In vitro studies showed that the Th1 clones were cytotoxic to λ2315 ‐transfected B lymphoma cells; by contrast, the Th2 clone was not cytotoxic in 5lCr‐release assay even though the Th2 cells inhibited the growth of λ2315 B lymphoma cells. The anti lympboma properties of both the Thl and Th2 clones appear to involve as yet undefined cytotoxic and growth inhibiting molecules.
Aberrant Tolerance Induction with Cationic Antigens
Authors: JACOBS M. J. M. , VAN DEN HOEK A. E. M. , VAN DE PUTTE L. B. A. , VAN DEN BERG W. B.
Published: July 2006

We studied the induction of tolerance in female C57B1/6 mice by oral and intravenous (i.v.) administration of protein antigens before immunization. Native proteins [chicken egg albumin (OVA), bovine serum albumin (BSA)] and their cationic derivatives [amidated (a)OVA, aBSA, methylated (m)BSA] were compared in their capacity to suppress cell‐mediated immunity (CMI), as measured by a delayed type hypersensitivity test (DTH). Oral feeding of 0.5 mg negative proteins gave a clear suppression. By contrast, cationic derivatives (50 μg to 50 mg) did not suppress CMI. Data from cross experiments, where aOVA was fed in OVA‐immune mice, showed no suppression at all. When OVA was fed in aOVA‐immune mice, only a partial suppression was achieved.

The potency to induce tolerance differed also when the antigens were administered i.v.: 25 μg OVA was sufficient to induce a clear suppression, whereas a much higher amount ofaOVA (500 μg) caused marginal suppression in OVA‐ and aOVA‐immune mice, respectively. Nevertheless, when concentrations of aOVA up to 2.5 mg were tested in a chronic model of arthritis, a significant suppression was achieved.

The differences in inducing a CMI suppression may have implications for the feasibility of immunointervention. Successful modulation of human arthritis may be highly dependent on the nature of the antigen involved.
Idiotypic Vaccination: Immunoprotection Mediated by Anti‐idiotypic Antibodies with Antibiotic Activity
Authors: POLONELLI L. , LORENZINI R. , DE BERNARDIS F. , GERLONI M. , CONTI S. , MORACE G. , MAGLIANI W. , CHEZZI C.
Published: July 2006

Anti‐Id antibodies were raised in mice against a monoclonal antibody (MoAb KT4) that neutralized the in vitro activity of a Pichia anomala yeast killer toxin. Monoclonal antibody was administered to BALB/C syngeneic mice with different schedules of immunization before intravenous challenge with increasing amounts of yeast killer toxin‐sensitive Candida alhicans cells. The course of candidosis was studied in comparison with mice non‐immunized and immunized with an isotypc‐matched unrelated MoAb subdivided into control groups. Protection was reflected by statistically significant increases in survival rate of mice immunized with MoAb KT4 which showed variable serum levels of yeast killer toxin‐like anti‐Id antibodies. MoAb KT4 affinity chromatography purified mouse anti‐Id antibodies were capable of killing in vitro the yeast ceils of the Candida albicans strain used for the experimental infection.

This is the first report of antimicrobial protection that exploits the role of anti‐idiotypic antibodies presumably acting in vivo as antibiotics (idiotypic vaccination).
Tumour‐Like Localized Amyloid of the Brain is derived from Immunoglobulin Light Chain
Authors: ERIKSSON L. , SLETTEN K. , BENSON L. , WESTERMARK P.
Published: July 2006

A patient is presented in whom the amyloid component of an intracerebral‘amyloidoma’has been purified and characterized by amino acid sequence analysis. The material originated from an autopsy of a 76‐year‐old man who 15 years earlier had been operated for an intracerebral‘amyloid tumour'. The tumour had recurred and grown to an almost walnut‐sized mass in the right cerebral hemisphere. It was located in the parietal lobe close to the lateral ventricle and had a close connection to the choroid plexus. Histological examination showed large masses of amyloid surrounded by some plasma cells and a few macrophages of the foreign body type, Amino acid sequence analysis of a major fibril subunit protein showed homology with the variable region of a monoclonal lambda immunoglobulin light chain, subgroup III or IV. This shows that the amyloid in the ‘tumour’ was of AL type and presumably derived from local synthesis by plasma cells.
Utilization of the MHC Class I (H‐2Kb) Purified Molecule and its Synthetic Peptides for Inhibition of Kb‐Specific Suppressor T Cells and their Induction In Vivo by the MHC Peptides
Authors: BRONDZ B. D. , ANFALOVA T. V. , PAVLOVA L. S. , PANKRATOVA E. V. , KOJICH A. G. , MOSHNIKOV S. A.
Published: July 2006

Six synthetic peptides of the MHC class I molecule corresponding to individual H‐2Kb participants in amino acid sequences of domains α1 (peptide 1 and 2) and α2 (peptides 3,4, 5,6) were selected. Kb‐specific suppressor T cells (Ts) were induced in vivo in mice, then pretreated with a set of peptides and assayed by proliferation decrease in a three‐cell lymphocyte culture (MLC). The effector function of Ts was abolished by the complex of the α2‐domain peptides (but nol by the α1‐domain peptides) and decreased by particular peptides separately (4, 5, 6) of the α2‐domain. Both α1‐ and α2‐domain peptides. added in high concentration, decreased otherwise efficient enrichment of Ts during the absorption‐elution procedure on the syngeneie macrophage (Mφ) monolayers. A similar significant effect was observed using the purified Kb molecule (100μg/ml) in the allogeneic Mφ monolayer. Interaction between Ts receptors and some MHC peptides indicates in effector Ts activation in vivo by induction with peptides 5 and 6 of the α2‐domain. The fine mechanisms of interaction between MHC class I molecule epitopes and T‐cell receptors of each of the T‐cell subsets separately are presently being studied.
In Vitro and In Vivo Measurement of Cell‐Mediated Immunity in Patients with HIV‐1 Infection
Authors: BORLEFFS J. C. C. , SCHUURMAN H.‐J. , VREHEN H. M. , SCHAIK M. VAN , BAST E. J. E. G.
Published: July 2006

We studied the usefulness of the in vitro lymphoproliferation assay and the in vivo skin lest in HIV‐1‐infected patients by using Chotridium tetani and tuberculin as testing antigens. Moreover, the relationship between data obtained from both assays was studied.

In 56 HIV‐infected patients not receiving antiretroviral therapy CD4+ cell counting was performed. In addition, in vitro (lymphocyte proliferation assay) and in vivo (delayed type hypersensitivity skin test) measuring of the immune status was done using C. tetani atid tuberculin as testing antigens.

When using C. tetani a significant correlation between the results of both tests and the CD4+ cell count was found. In contrast to earlier reports from African countries, in vivo skin testing using tuberculin did not yield clinically significant information on the degree of Immunodeficiency. We explain our findings by the fact that healthcare policy in The Netherlands encompasses vaccination with C. tetani. which enables the application of C. tetani as testing antigen for measuring immune function both in vitro and in vivo.
The Structure of the Variable Regions of Mouse Monoclonal Antibodies to Hepatitis B Virus Core Antigen
Authors: SKRIVELIS V. , STEINBERG Yu. , BICHKO V. , GREN E. , TSIMANIS A.
Published: July 2006

From a panel of monoclonal antibodies (MoAbs) directed against E. coli‐derived native and denatured hepatitis B virus (HBV) core antigen we have selected a set of specific MoAbs which recognize different linear antigenic determinants: MoAb C1‐5—cl epitope; MoAb 14K8—less immunogenic N‐terminal region; and MoAbs 13C9, 10F10 and 14E11, 14G3—the immunodominant region between amino acids 134 and 140.

We have applied the polymerase chain reaction technique to clone Ig VH and YL, region genes, and appropriate full‐length cDNA clones were obtained and characterized by nucleotide sequence analysis. Among the six heavy chain variable region sequences examined, three Yh families were represented. Two of them belong to the 7183 (MoAb Cl‐5) and 3609 (14B8) families respectively and four, having only two amino acid changes in the CDR2 region, to the J558 family. These four probably are derived from a single expanded B‐cell clone. The light chain sequences indicate that their YL are encoded by YK21, VK19 and YK3 germline genes. Unlike YH genes, light chain genes are closely related to known representatives of mouse kappa light chain families and are employed also by MoAbs raised against other antigens.
Antibody Response to Meningococcal Polysaccharides A and C in Patients with Complement Defects
Authors: BISELLI R. , CASAPOLLO I. , D'AMELIO R. , SALVATO S. , MATRICARDI P. M. , BRAI M.
Published: July 2006

Patients with defects of terminal complement components are particularly exposed to the risk of developing neisserial infections and seem to respond poorly to meningococcal capsular polysaccharide (PS) C via natural immunization. The sole meningococcal PSC is. on the other hand, an excellent immunogen in normal people. Considering the great importance of vaccine prophylaxis for the prevention of meningococcal infections in patients with complement defects, it is crucial to study the antibody response to the sole meningococcal PS in these patients. We therefore analysed the levels of anti‐PSA and PSC antibodies in the members of four families including patients with homozygous and heterozygous defects of C7, C8 or factor H, before and after vaccination with the sole PSA+C.

Surprisingly, we found the highest levels of antibodies before vaccination In homozygous subjects, followed by heterozygous and normal controls, whereas, after vaccination, homozygous subjects showed the lowest increase of specific antibodies, indicating their relative incapability to respond to sole meningococcal PS.

In conclusion, this study demonstrates (1) the capacity to respond to meningococcal PS via natural immunization by patients with total complement defects, and (2) the low responsiveness to meningococcal PS via vaccine immunization by the same patients. We propose that vaccination should be given to patients lacking specific antibodies and their serological response should be assessed. In addition this study confirms previous observations on a likely lower immunogenic power of meningococcal serogroup C via natural immunization compared with the better immunogenicity of the sole PSC.
Cellular Interactions between Synovial T‐Cell Clones from a Patient with Rheumatoid Arthritis
Authors: LAAR J. M. VAN , MILTENBURG A. M. M. , KUIPER P. DE , DAHA M. R. , VRIES R. R. P. DE , BREEDVELD F. C.
Published: July 2006

The aim of this study was to investigate T‐T cell interactions that may be involved in the pathogenesis of chronic synovitis. Two CD4+ T‐cell clones, termed N16 and N18, sharing similar cytokine profiles were isolated from the synovial tissue of a patient with rheumatoid arthritis. Activated N16 cells induced proliferation of N18 cells in the absence of accessory cells, while the proliferative response of N18 cells to activated N18 cells depended on the presence of accessory cells. Whereas the response of N18 to activated N18 cells could be blocked by MoAbs against HLA class I and LFA‐1 molecules, the response of N18 to activated N16 cells was only weakly affected, suggesting that the surface molecules involved in this interaction differed. Indeed, using MoAbs against CD3, CD4 and to a lesser extent CD2, proliferation of N18 to activated N16 cells was abrogated. These data provide a model for T‐T cell interactions involved in the development of chronic synovitis.
Cell‐Mediated Immune Responses in Renal Transplant Recipients treated with Cyclosporin and Prednisolone with or without Azathioprine
Authors: WRAMNER L. , MJÖRNSTEDT L. , RYDBERG L. , OLAUSSON M.
Published: July 2006

The cell‐mediated immune response was studied, using mixed lymphocyte reactivity (MLR) and cell‐mediated lympholysis (CML) tests, in patients with well‐functioning kidney grafts from living donors at 6 and 12 months and at 2–5 years after transplantation. The patients were allocated to treatment with cyclosporin A (CyA) and prednisolone (group A) or with CyA, prednisolone and azathioprine (group B). The MLR towards a third party were in the range of that of untreated controls while the anti‐donor activities were reduced after 6 months in both groups. The CML activities in group A towards a third party were in the range of that of untreated controls at all times, while the anti‐donor activities were decreased. By contrast, the CML activities in group B towards a third party were decreased during the first year and were in the range of that of normal controls at 2–5 years. The anti‐donor CML activities were low at all times in group B.

In summary, the two CyA protocols allowed the induction of donor‐specific unresponsiveness within the first post‐transplant year. The anti‐third party activities were low during the first post‐transplant year in recipients with triple therapy but not in those without azathioprine.
Human Monoclonal IgG Rheumatoid Factors from the Synovial Tissue of Patients with Rheumatoid Arthritis
Authors: RANDEN I. , THOMPSON K. M. , THORPE S. J. , FØRRE Ø. , NATVIG J. B.
Published: July 2006

We have established IgG rheumatoid factor (RF)‐secreting hybridoma cell lines from the synovial tissues of two patients (TS and SJ) with rheumatoid arthritis (RA) und one (KL) with the polyarlicular form of juvenile rheumatoid arthritis (JRA). The IgG RF bind human and rubbit IgG and all except one form intracellular complement‐fixing complexes indicative of a self‐association process. The possibility of IgG RF for self association and immune complex formation is a feature thought to be important for the inflammatory processes in RA. Of the IgG RF‐secretingcell lines established, three clones from patient SJ and one from patient KL are of the IgG Ik isotype while five clones from patient TS are of the IgG2λ isotype.
T‐Cell Receptor Alpha Haplotype influences Vα Epitope Expression on both Cortisone‐Resistant Thymocytes and Lymph Node T Cells
Authors: BOGEN B. , GLEDITSCH L. , TEIG A.
Published: July 2006

We have previously described a mouse strain congenic for the T‐cell receptor alpha chain locus (Tcra). The strain, B10.D2.C‐Tcraa/Bo, carries the Tcraa haplotype of BALB/c instead of the Tcraa haplotype normally found in B10.D2. By comparing B10.D2.C‐Tcraa and B10.D2 mice, we now show that the Tcra haplotype influences the frequencies of Vα8 epitope expression on T cells detected by the novel B21.14 monoclonal antibody (MoAb). This finding agrees well with our previous observation that Tcra haplotype influences the frequencies of expression of two other Vα8 epitopes, detected by the KT50 and KT65 MoAbs. The specificities of the three Vα8‐specific MoAbs were compared by scatter diagram analysis of staining frequencies obtained in individual mice. The B21.14 and KT50 MoAbs appear to have very similar specificities; these two MoAbs stain slightly more αβ T cells than the KT65 MoAb. Investigations with another novel MoAb, the Vα2‐specific B20.1, revealed that the Tcra haplotype also influences the frequency of Vα2 epitope expression. The effect of the Tcra haplotype on Vα2 epitope expression was evident not only among lymph node T cells but also among mature, cortisone‐resistant thymocytes. Thus, the influence of the Tcra haplotype is imprinted already in the thymus. The most likely explanations for Tcra haplotype dependency of Vα epitope expression are differing numbers of Vα‐gene segment subfamily members and/or allelic polymorphism. The significance of Tcra haplotype‐dependent Vα epitope expression is discussed.
Macrophage Populations in Different Stages of Induced Hepatic Metastases in Rats: an Immunohistochemical Analysis
Authors: HEUFF G. , VAN DER ENDE M. B. , BOUTKAN H. , PREVOO W. , BAYON L. G. , FLEUREN G. J. , BEELEN R. H. J. , MEIJER S. , DIJKSTRA C. D.
Published: July 2006

Macrophages play a role in the host defence against cancer. Little is known about changes in macrophage populations during early metastatic growth. To evaluate the distribution, number and phenotype of macrophages in the development of hepatic metastases in a rat model (Wag/Rij rats and syngeneic CC531 colon carcinoma cell line), an immunohistochemical study was performed with the monoclonal antibodies ED1 (monocytes, and all macrophages), ED2 (resident tissue macrophages, like Kupffer cells) and ED3 (a subpopulation of macrophages which may play a role in the recruitment of lymphocytes). OX19 and Hisl4 were used to identify lymphocytes. In this study a new monoclonal antibody CC52 is described, which recognizes the CC531 tumour cell line. Liver metastases were induced by injection of CC53I colon carcinoma cells into a mesenteric vein. Rats were killed at various intervals. Results show three major macrophage populations during hepatic tumour growth: (1) on day 3, infiltrates are observed around the micrometastases, which contain mainly newly recruited macrophages (ED1+ and ED2−); (2) after 7 days, ED3‐positive (ED3 +) macrophages together with T lymphocytes are found in the infiltrates; (3) an increase in the number of ED2‐positive (ED2+) Kupffer cells is observed in the liver parenchyma after 14 days. In conclusion, the present results suggest that various populations of macrophages, newly recruited (ED1+) as well as resident Kupffer cells (ED2+), are involved in the immune response against tumour cell deposits in the liver.
Antibody Response to Pneumococcal Polysaccharide Vaccine in Young, Adult and Old Mice
Authors: AABERGE I. S. , NORTH R. J. , GROENG E.‐C. , LØVIK M.
Published: July 2006

The anti‐pneumococcal antibody response was studied in young (5‐week‐old) and adult (10‐week‐old) BALB/c and CBA/J mice and in adult (9–10‐week‐old) and old (12‐, 18‐ and 24‐month‐old) AB6F1 and B6D2F1 mice after s.c. immunization with a 23‐valent pneumococcal polysaccharide vaccine. Both young and adult mice showed a significant IgM antibody response to the vaccine 6 days after immunization with 111 /ig antigen. There were significant immune responses to serotypes 1, 2, 4 and 7F in contrast to small responses to serotypes 14, 19F and 23F after immunization with the vaccine. One month after immunization, there were only marginal differences in IgM anti‐pneumococcal antibody levels to the vaccine (anti‐PPS) between immunized and unimmunized BALB/c mice, whereas in CBA/J mice the anti‐PPS remained higher in immunized than in unimmunized mice. Immunization of old mice induced a significant IgM antibody response 6 days after immunization, but the anti‐PPS thereafter decreased rapidly towards preimmunization values in AB6F1 mice. A significant IgG anti‐PPS was not detected in any of the mice studied. The IgA anti‐PPS tended to vary over time with no consistent pattern. It is important to carefully consider age and strain of the mice used when studying the immune response to pneumococcal polysaccharide antigens.