Variability in the biological response to anti-CD20 B cell depletion in systemic lupus erythaematosus.

OBJECTIVE: To study the effects in systemic lupus erythaematosus (SLE) of B cell directed therapy with rituximab, a chimeric monoclonal antibody directed at CD20+ B cells, without concomitant immunosuppressive therapy in mild to moderate SLE. METHODS: Patients (n=24) with active SLE and failure of >or=1 immunosuppressive were recruited from three university centres into this phase I/II prospective open-label study. Patients were followed for 1 year to assess safety, efficacy and biological effects. RESULTS: In total, 18 of the patients scheduled to receive the full lymphoma dose of rituximab were evaluable for B cell levels in peripheral blood. Of these, 17 had effective CD19+ B cell depletion (<5 cells/microl). However, six of the depleted patients showed B cell return before 24 weeks. A total of 70% of patients improved by week 55, as defined by an SLE Disease Activity Index (SLEDAI) score improvement of >or=2 units from baseline. The degree of CD19+ B cell depletion was correlated with SLEDAI improvement at week 15 (r=0.84). In general, rituximab infusions were well tolerated. Approximately a third of the patients developed human anti-chimeric antibody (HACA) titres, which correlated with poor B cell depletion. Most patients (9 of 14) did not respond to immunisations with Pneumovax and tetanus toxoid. CONCLUSIONS: Rituximab is a promising new therapy for SLE. The variability of responses in patients with SLE may be related to HACA formation. The failure to respond to immunisations is surprising, in view of the apparently low risk of infections. Better biological markers are necessary to follow these patients during treatment.

B cells as a therapeutic target in autoimmune diseases other than rheumatoid arthritis.

Selective B-cell depletion with anti-CD20 therapy is a promising novel treatment option for patients with refractory autoimmune disease. The anti-CD20 antibody, rituximab, is the first therapeutic monoclonal antibody to have been approved by the European Medical Agency (EMEA) and the US Food and Drug Administration (FDA) for the treatment of relapsed, low-grade, follicular non-Hodgkin's lymphoma. Rituximab is now being studied in a range of autoimmune diseases, most notably rheumatoid arthritis, but also chronic immune thrombocytopenic purpura and systemic lupus erythematosus. Current data obtained from studies of rituximab single-agent therapy for autoimmune disease show good tolerability and sustained improvement in disease symptoms, although the precise mechanisms of action in autoimmunity remain to be fully clarified. Future research is likely to be focused on the optimization of responses with rituximab-based therapy. However, early observations suggest that this approach is likely to yield significant clinical benefits in a wide range of organ-specific and systemic autoimmune diseases.

Panel discussion on B cells and rituximab: mechanistic aspects, efficacy and safety in rheumatoid arthritis and non-Hodgkin's lymphoma.

The clinical potential of rituximab (MabThera/Rituxan), a selective B-cell-depleting agent, in the treatment of patients with rheumatoid arthritis (RA) is rapidly becoming apparent. The data presented at an official satellite symposium of the European League Against Rheumatism (EULAR) Congress (2003, Lisbon, Portugal), reinforce the rationale for the use of this novel agent in RA and have provided an early indication of its clinical efficacy, safety and tolerability. The symposium presentations were followed by a panel discussion and a question and answer session in which the participants were able to shed further light on specific mechanistic issues relating to effects on B-cell populations based on available data and their own clinical experience of rituximab. Additionally, the implications of current results for longer-term clinical efficacy and safety were discussed. It is becoming clear that rituximab (alone or in combination with disease-modifying anti-rheumatic drugs) is highly efficacious in RA. Extensive data from patients with non-Hodgkin's lymphoma show that early concerns over increased infection rates due to prolonged suppression of B cells have not been realized. The effects of rituximab on long-term RA outcomes, such as joint erosion and duration of response (particularly in patients receiving combination therapy), are eagerly anticipated.

RANK, RANKL and OPG in inflammatory arthritis and periprosthetic osteolysis.

Elucidation of the receptor activator of nuclear factor kappa B (RANK), its ligand (RANKL) and osteoprotegerin (OPG) as the final effectors of bone resorption has transformed our understanding of metabolic bone diseases and revealed novel therapeutic targets. Activation of the RANK-RANKL signaling pathway is directly responsible for dramatic focal erosions that are observed in inflammatory arthritis and aseptic loosening of orthopaedic implants. While these conditions share many features common to all metabolic bone disorders (e.g., osteoclastic resorption), they exhibit several unique properties, which are highlighted in this review. Most important is the relative inability of bisphosphonate therapy to inhibit osteolysis in joint inflammation and periprosthetic joint loosening and the unexpected effectiveness of anti-cytokine therapy in both rheumatoid and psoriatic arthritis. Herein, we provide a review of the role of RANK, RANKL and OPG in erosive arthritis and periprosthetic osteolysis and discuss the potential of anti-RANKL therapy for these conditions.

B lymphocytes in systemic lupus erythematosus: lessons from therapy targeting B cells.

Systemic lupus erythematosus (SLE) is a complex disease characterized by numerous autoantibodies and clinical involvement in multiple organ systems. Autoantibodies are usually present in serum for years before the onset of clinical disease. Autoimmunity begins with a limited number of autoantibodies and evolves to become progressively more diverse. Eventually clinical disease ensues. The immunological events triggering the onset of clinical manifestations have not yet been defined. While undoubtedly T cells and dendritic cells appear to play major roles in SLE, a central role for B cells in the pathogenesis of this disease has been brought to the fore in the last few years by work performed both in mice and humans by multiple laboratories. As a result, there is little doubt about the importance of B cells in the development of SLE. Yet much remains to be learned about their role in the ongoing disease process and the merit of targeting B cells for the treatment of SLE. This article will review the role of B cells in human SLE as well as the currently available data on the treatment of SLE by depleting B cells with anti-CD20 (rituximab).

Oral pentoxifylline inhibits release of tumor necrosis factor-alpha from human peripheral blood monocytes : a potential treatment for aseptic loosening of total joint components.

BACKGROUND: Pentoxifylline (Trental) is a methylxanthine-derivative drug that has been used for more than twenty years in the treatment of peripheral vascular disease. Pentoxifylline is also a potent inhibitor of tumor necrosis factor-alpha (TNF-alpha) secretion, both in vitro and in vivo, and has demonstrated efficacy in the treatment of certain animal and human inflammatory diseases. Pentoxifylline has a potential therapeutic role in the treatment of aseptic loosening of total joint replacement components because it inhibits TNF-alpha secretion by particle-stimulated human peripheral blood monocytes. The purpose of our study was to determine whether the particle-stimulated secretion of TNF-alpha by peripheral blood monocytes was inhibited in volunteers who had received pentoxifylline orally. METHODS: Human peripheral blood monocytes were harvested from eight healthy volunteers and were exposed to three different concentrations of titanium particles or to 500 ng/mL of lipopolysaccharide as a positive control. The same volunteers were then given pentoxifylline (400 mg, five times per day) for seven days. Their peripheral blood monocytes were again isolated and exposed to experimental conditions, and the TNF-alpha levels were measured. RESULTS: The peripheral blood monocytes from all eight volunteers showed a significant reduction in TNF-alpha release following oral treatment with pentoxifylline. This reduction was observed at exposures of 10(7) and 10(6) titanium particles/mL and in the lipopolysaccharide-treated group, but not at 10(5) particles/mL. CONCLUSIONS: To our knowledge, this is the first study to demonstrate the ability of an oral drug to decrease the release of TNF-alpha from human peripheral blood monocytes exposed ex vivo to particle debris. TNF-alpha is involved in the pathogenesis of osteolysis and subsequent loosening of total joint arthroplasty components. The ability to suppress the release of TNF-alpha in patients with a total joint replacement may help to control osteolysis and to reduce the development of aseptic loosening. This effect could increase implant longevity and decrease the need for revision arthroplasty.

Hepatitis B immunization of healthy elderly adults: relationship between naïve CD4+ T cells and primary immune response and evaluation of GM-CSF as an adjuvant.

The efficacy of granulocyte-macrophage colony-stimulating factor (GM-CSF) to enhance the primary immune response to hepatitis B vaccine was studied in healthy elderly with young volunteers included as controls in this double-blind, placebo-controlled trial of GM-CSF as an immune adjuvant. Naïve T-helper cells (CD4+CD45RA+) were determined at baseline. Forty-five healthy elderly (average age, 74 years) and 37 healthy young controls (average age, 28 years) were randomized. Hepatitis B vaccine was administered at 0, 1, and 6 months. GM-CSF as a single injection of either 80 microg or 250 microg with the first and second doses of hepatitis B vaccine. In this trial GM-CSF did not enhance antibody responses. However, the antibody responses were dramatically different between these two groups: 35/35 young developed a protective titer versus 19/45 elderly (P < 0.0001). In addition, the mean logarithm of anti-hepatitis B antibody level in the 35 young who completed the study was 3.17 (log mIU/ml) but only 2.21 in the 19 elderly responders (P < 0.0001). Naïve T-helper cells differed significantly between the two groups: the mean percentage of CD4+CD45RA+ T cells was 47.9% versus 35.0% (P < 0.0001) in the young and elderly volunteers respectively. Naïve T cells also differed significantly between elderly who did or did not respond to HBV (39.9% vs. 31.7%, P = 0.039). Using linear regression, age, and percent naive, CD4 T cells were determined to significantly influence the anti-hepatitis B antibody response, but sex and dose of GM-CSF did not. For a two-parameter model: logarithm of antibody titer = (-0.038 x age in years) + (0.031 x % naïve CD4T cells) + 2.68; adjusted r2 = 0.605 and P < 0.0001. However, age had a larger effect than naive CD4 T cells, i.e., in comparing young and elderly groups the log antibody titer decreased by 1.73 due to the increase in age but only 0.40 due to the decrease in naive CD4 T cells. Thus, there was a large effect of age that could not be explained by the quantitative change in the naïve T-helper cells.

Response of C-reactive protein and serum amyloid A to influenza A infection in older adults.

Influenza epidemics are associated with significant morbidity and mortality in the elderly, with a substantial proportion of deaths due to cardiovascular events. Elevations of acute-phase proteins have been associated with an increased risk of atherosclerotic events. Therefore, serum amyloid A (SAA) and C-reactive protein (CRP) were measured during influenza illness and 4 weeks later in 7 young persons, 15 elderly outpatients, and 36 hospitalized adults. Striking elevations were seen in mean acute SAA and CRP levels in all groups, but hospitalized patients had the highest levels (SAA, 503 vs. 310 microg/mL [P=.006]; CRP, 120 vs. 34 microg/mL [P<.001]). The presence of dyspnea, wheezing, and fever was also associated with high CRP levels. Influenza infection is associated with significant elevations of SAA and CRP levels in elderly patients, especially those who require hospitalization. It is possible that direct effects of CRP may exacerbate preexisting atherosclerotic lesions and may help explain cardiovascular events associated with acute influenza.

Anti-TNF-alpha therapy as a clinical intervention for periprosthetic osteolysis.

Aseptic loosening of total joint arthroplastics due to periprosthetic osteolysis is a frequent cause of implant failure. The absence of clinical interventions to arrest or prevent this complication limits the use of total joint replacement especially in younger patients. Here we review recent studies implicating tumor necrosis factor (TNF)-alpha in periprosthetic osteolysis and the rationale for clinical studies of anti-TNF therapy and other interventions for periprosthetic loosening.

Comparison of respiratory syncytial virus humoral immunity and response to infection in young and elderly adults.

Little information about immunity to respiratory syncytial virus (RSV) and disease pathogenesis in elderly persons exists. Humoral immunity to RSV was assessed in 41 young, 56 healthy elderly, and 49 frail elderly adults by measuring baseline RSV specific IgG by enzyme immunoassay (EIA) and microneutralization assay (MNA) in serum. A comparison of the immune response of 11 young and 28 elderly persons with natural RSV infection was also performed. Despite significant differences in age and functional status, no decreases in RSV antibody levels by either EIA or MNA were noted in the elderly compared with the young. Mean baseline MNA titers expressed as log2 were 10.5 +/- 1.1 for the young, 10.5 +/- 1.5 for the healthy elderly, and 10.9 +/- 1.6 for the frail elderly. The frail elderly who attend a daycare had the highest RSV titers to F by EIA at 16.6 +/- 2.0, compared with 15.4 +/- 1.4 and 15.1 +/- 1.4 in the healthy elderly and young, respectively. This finding may reflect recent infection due to their communal setting or increased production of non-neutralizing antibody. The immune response of older persons to RSV infection was as vigorous as the younger subjects, with 79% having a >/=fourfold rise in MNA titers compared to 64% in the young. These data suggest that the severe clinical manifestations of RSV in the elderly are not due to a significant defect in humoral immunity.

Cytomegalovirus seropositivity is associated with the expansion of CD4+CD28- and CD8+CD28- T cells in rheumatoid arthritis.

OBJECTIVE: Previous researchers have found expansion of CD4+CD28- T cells in patients with rheumatoid arthritis (RA) compared to age matched controls, and have identified expanded clones of autoreactive cells within this population. We examine the association of prior cytomegalovirus (CMV) infection (positive serum anti-CMV IgG) with the percentage of CD4+CD28- T cells and CD8+CD28- T cells in patients with RA. METHODS: A total of 45 patients (36 women, 9 men), mean age of 59 years, with definite RA were studied. RESULTS: In this group 28 patients were seropositive for CMV and 17 seronegative. Seropositive and seronegative subjects did not differ significantly in age, sex, medication use, or severity of disease. Joint count, Health Assessment Questionnaire, pain score, patient global assessment, physician global assessment, and presence of extraarticular disease served to assess disease severity. Expression of CD4/CD28/CD57 and CD8/CD28/CD57 on lymphocytes was determined by 3 color flow cytometry. (CD28 and CD57 are reciprocally related.) CD4+CD28-CD57+ T cells were expanded only in CMV seropositive patients. CONCLUSION: The "carrier" phenotype that has been hypothesized based on a 2 population model for the distribution of CD4+CD28- T cells in RA can be explained by prior infection with CMV.

Role of cytomegalovirus in the T cell changes seen in elderly individuals.

The effect of prior cytomegalovirus (CMV) infection on the immune system was evaluated in young and elderly volunteers. Prevalence of IgG antibodies to CMV was higher in the elderly volunteers. In both age groups, there was a strong association with CMV seropositivity and increased number of CD28- CD4 or CD8 T cells, as well as with increased numbers of T cells expressing CD56 or DR. Although these changes have previously been reported to be age-related, they were independent of age when CMV serological status was taken into account. In contrast, both age group and CMV status were important determinants of the total number of T cells, the number of CD8 T cells, and the number of CD8 T cells expressing CD45RA or CD28. These findings indicate that prior infection with CMV, as reflected by CMV serological status, has important effects on T cell subsets and surface markers and must be considered whenever evaluating age-related changes in immunological parameters.

Acute respiratory exposure of human volunteers to octamethylcyclotetrasiloxane (D4): absence of immunological effects.

Humans are exposed to silicones in a number of commercial and consumer products. Some of these silicones, including octamethylcyclotetrasiloxane (D4), are volatile. Therefore, there is a potential for respiratory exposure. A pharmacokinetic analysis of respiratory exposure to D4 is presented in the accompanying paper (M. J. Utell et al., 1998, Toxicol. Sci. 44, 206-213). Possible immune effects of respiratory exposure to D4 are investigated in this paper. Normal volunteers were exposed to 10 ppm D4 or air for 1 h via a mouthpiece using a double-blind, crossover study design. Assays were chosen to screen for immunotoxicity or a systemic inflammatory response. Assessment of immunotoxicity included enumeration of peripheral lymphocyte subsets and functional assays using peripheral blood mononuclear cells. Because in humans there is no direct test for adjuvant effect of respiratory exposure, we analyzed proinflammatory cytokines and acute-phase reactants in peripheral blood, markers for a systemic inflammatory response, as surrogate markers for adjuvancy. These tests were repeated when the volunteers were reexposed to D4 approximately 3 months after this initial exposure. Blood was obtained prior to exposure, immediately postexposure, and 6 and 24 h postexposure. In these short-term, controlled human exposures, no immunotoxic or proinflammatory effects of respiratory exposure to D4 were found.

Patterns of cytokine production in psoriatic synovium.

OBJECTIVE: To determine patterns of cytokine production and mRNA expression in synovium from patients with psoriatic arthritis (PsA) and to compare the profile of cytokine production in PsA explants with those derived from rheumatoid (RA) and osteoarthritic (OA) synovia and psoriatic skin. METHODS: Cytokine levels were measured in supernatants from synovial and dermal explant cultures at Day 10 by ELISA. Cytokine mRNA expression in PsA whole tissue was determined by multi-gene assay. Cytokine levels in explant supernatants were compared between PsA, RA and OA, and psoriatic skin. Synovial tissues were scored histologically by a pathologist blinded to the clinical diagnosis. RESULTS: PsA explants released elevated levels of interleukin (IL)-1beta, IL-2, IL-10, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha, but not IL-4 or IL-5. A similar pattern of gene expression was detected in whole synovial tissue. These cytokine levels were greater in PsA than RA, despite higher histopathologic scores in RA explants. Production of IL-1beta, IFN-gamma, and IL-10 were strongly correlated. Levels of IFN-gamma, IL-1beta, and IL-10 were higher in psoriatic synovium than psoriatic dermal plaques. CONCLUSION: The cytokine profile in PsA is characterized by the presence of Th1 cytokines and the monokines TNF-alpha and IL-1beta and very elevated levels of IL-10. The higher levels of these cytokines in PsA compared to RA suggest the presence of different underlying mechanisms.

Airway inflammation in smokers and nonsmokers with varying responsiveness to ozone.

Exposure to ozone causes symptoms, changes in lung function, and airway inflammation. We studied whether individuals who differ in lung-function responsiveness to ozone, or in smoking status, also differ in susceptibility to airway inflammation. Healthy subjects were selected on the basis of responsiveness to a classifying exposure to 0.22 ppm ozone for 4 h with exercise (responders, with a decrease in FEV1 > 15%; and non-responders, with a decrease in FEV1 < 5%). Three groups were studied: nonsmoker-nonresponders (n = 12), nonsmoker-responders (n = 13), and smokers (n = 13, 11 nonresponders and two responders). Each subject underwent two exposures to ozone and one to air, separated by at least 3 wk; bronchoalveolar and nasal lavages were performed on three occasions: immediately (early) and 18 h (late) after ozone exposure, and either early or late after air exposure. Recovery of polymorphonuclear leukocytes (PMN) increased progressively in all groups, and by up to 6-fold late after ozone exposure. Interleukin-6 (IL-6) and IL-8 increased early (by up to 10-fold and up to 2-fold, respectively), and correlated with the late increase in PMN. Lymphocytes, mast cells, and eosinophils also increased late after exposure. We conclude that ozone-induced airway inflammation is independent of smoking status or airway responsiveness to ozone.

Tissue-specific expression of the human prostate-specific antigen gene in transgenic mice: implications for tolerance and immunotherapy.

Human prostate-specific antigen (PSA) has been widely used as a serum marker for cancer of the prostate. The cell type-specific expression of PSA also makes it a potential tumor antigen for prostate cancer immunotherapy. Study of the immunological aspects of PSA within either normal or malignant prostate tissue has been hampered by the lack of a mouse model, because no PSA counterpart has been identified in mice. Using a 14-kb genomic DNA region that encompasses the entire human PSA gene and adjacent flanking sequences, we generated a series of human PSA transgenic mice. In the six independent lines of transgenic mice generated, the expression of the human PSA transgene, driven by its own cis-acting regulatory elements, is specifically targeted to the prostate. Tissue distribution analysis demonstrated that PSA transgene expression closely follows the human expression pattern. Immunohistochemical analysis of the prostate tissue also showed that the expression of the PSA transgene is confined to the ductal epithelial cells. Despite expressing PSA as a self-antigen in the prostate, these transgenic mice were able to mount a cytotoxic immune response against PSA expressed by tumor cells, indicating that expression of the transgene has not resulted in complete nonresponsiveness. This transgenic mouse model will provide a well defined system to gain an insight into the mechanisms of nonresponsiveness to PSA, ultimately leading to strategies for immunotherapy of human prostate cancer using PSA as the target antigen.

Regulation of IgE and cytokine production by cAMP: implications for extrinsic asthma.

Recent investigations indicate that pharmacologic agents which elevate intracellular levels of cyclic AMP (cAMP) also enhance immunoglobulin E (IgE) production. This review proposes that elevation of intracellular cAMP is a prominent mechanism which enhances IgE production. Enhancement is mediated by two mechanisms. First, cAMP-elevating agents directly target B lymphocytes, promoting recombination of the Ig heavy chain loci. Second, these agents indirectly promote IgE production by inducing a T-helper type 2 (Th2) profile of cytokine secretion. In turn, Th2-type cytokines interact with B lymphocytes and direct isotype switching to the epsilon locus. One type of cAMP-elevating agents, the beta2-adrenergic receptor agonists (beta2-agonists), are used to treat asthma. A number of detrimental phenomena have been associated with beta2-agonist use such as, rebound hyperresponsiveness and increases in asthma mortality. This review theorizes that beta2-agonists enhance IgE and Th2 cytokine production and that these mediators exacerbate extrinsic, IgE-dependent asthma.