Local Antibody Production Research Articles

Oral immunization induces local and distant mucosal immunity in swine

Transmission of porcine reproductive and respiratory syndrome virus (PRRSV) in semen and reproductive disease in pregnant swine might be reduced by vaccines that induce mucosal immunity in the reproductive tract. Cholera toxin (CT), when delivered orally, is a potent mucosal adjuvant and immunogen in swine. To determine if oral immunization additionally elicits immunity at distant mucosal surfaces, we examined antibody responses to CT-B subunit in the reproductive tract and oral cavity. Orally administered CT induced distant mucosal immunity, as measured by antibodies to CT-B subunit in saliva and vaginal secretions. Presentation of PRRSV nucleocapsid as a genetic fusion with CT resulted in local mucosal antibody production, but no response was observed in vaginal secretions. The results demonstrate the feasibility of using orally administered CT for the induction of immunity to reproductive pathogens in swine. However, effective induction of PRRSV-specific immune responses in the reproductive tract requires a better understanding of the mechanisms of antigenicity and adjuvanticity at distant mucosal sites.

Aqueous humor and serum immunoblotting for immunoglobulin types G, A, M, and E in cases of human ocular toxoplasmosis.

The purpose of this study was to compare the local and systemic Toxoplasma-specific humoral immune responses in individuals with ocular toxoplasmosis (OT). To this end, paired aqueous humor and serum samples from 46 individuals with active OT and from 30 individuals without inflammatory eye disease (controls) were analyzed by immunoblotting for anti-Toxoplasma immunoglobulin G (IgG), IgA, IgM, and IgE directed against 20- to 120-kDa antigens. The presence in the aqueous humor of a unique band, or of at least three bands that were at least three times more intense in aqueous humor than in serum, was taken as evidence of local antibody production. IgG bands were detected in 98% of the aqueous humor samples, while IgA bands were detected in 76%, IgM bands were detected in 8%, and IgE bands were not detected in any. Evidence of local production of specific antibodies was found in 32 cases (70%) (IgG in 23 [50%]; IgA in 16 [35%]). In 10 instances (22%), routine laboratory tests were not indicative of OT. In 14 cases (30%), no local antibody production was detected by immunoblotting; 3 of these cases yielded evidence of local antibody production according to the Goldmann-Witmer coefficient. Local antibody production was revealed for 7 of the 30 controls (23%). Hence, the sensitivity of immunoblotting for IgG and IgA is 70%, and the specificity is 77%. We conclude that immunoblotting for local specific IgG and IgA supports the clinical diagnosis of OT in 70% of cases. In 22% of these, the diagnosis is not confirmed by other laboratory tests. Hence, immunoblotting increases the sensitivity of routine laboratory tests and should be considered for samples that register negative by such tests.

Use of fluorescence resonance energy transfer hybridization probes to evaluate quantitative real-time PCR for diagnosis of ocular toxoplasmosis.

Toxoplasma gondii infection is an important cause of chorioretinitis in Europe and the United States. Ophthalmological examination and a good clinical response to adequate therapy mainly support ocular toxoplasmosis diagnosis. However, clinical diagnostic may be difficult in some atypical cases. In these cases, laboratory confirmation, based on detection of local specific antibodies and parasite DNA by conventional PCR, is therefore important to confirm the disease etiology. More recently, real-time PCR has been developed to improve prenatal congenital toxoplasmosis diagnosis. We therefore examined the diagnostic value of quantitative real-time PCR for the detection of T. gondii in aqueous humor samples, associated with quantification of human beta-globin to control sample quantitative quality, by using a double fluorescence resonance energy transfer hybridization probes system with a double fluorescence reading. Of the 23 the clinically toxoplasmosis suspect patients, 22 showed serological evidence of exposure to Toxoplasma; one had a serological profile indicative of active infection. The analysis of paired aqueous humor and serum samples revealed an intraocular antibody production in 9 of 23 cases (39.1%). The quantitative real-time PCR revealed positive and high parasite numbers and high Toxoplasma/human genome ratios in three cases. Furthermore, PCR was the only positive confirmatory test in two cases (11.1%). None of the patients included in the control group (n = 7) had evidence of either local specific antibody production or T. gondii DNA detection, suggesting a good relative assay specificity. On the whole, quantitative real-time PCR appears to be useful for diagnosing atypical ocular toxoplasmosis presentations.

Actualités cliniques et diagnostiques de l’herpès cornéen

The indications for keratoplasty in treating herpes keratitis are currently declining because of recent progress in diagnosis and treatment. Clinically, corneal signs may be caused by HSV reactivation or a secondary anti-HSV immune response. Corneal opacification may be acute or the expression of sequela (meta-herpetic keratitis). The virus can be detected on a corneal surface sample by direct examination or cell culture, the only way to detect an infective virus. The detection of local antibody production in the aqueous humor is an inexpensive method, indicating the local immune anti-HSV response. Detection of HSV DNA using PCR is more sensitive, but the presence of HSV DNA within corneal tIssue may be more delicate to interpret. It is now proven that HSV can be transmitted through a corneal graft from donor to recipient, but no diagnostic test currently detects potentially infective corneas in eyebanks.

Evidence for local production of antibodies to auto and non-self antigens in periodontal disease.

Autoimmune mechanisms may contribute to periodontal disease (PD) pathogenesis; autoantibody to collagen type 1 is produced at the periodontal site and local levels are found to be higher than in serum. To find any evidence of autoimmune destruction in diseased periodontal tissues in patients with periodontitis. The study examines the relationship of antibodies to a self antigen collagen Type 1 and antigens from two periodontal pathogens namely Porphyromonas gingivalis (Pg), Actinobacillus actinomycetemcomitans (Aa) and a non-oral bacterium Bacteroides fragilis (Bf) in disease sites and in serum. Granulomatous tissues from periodontally diseased sites and serum samples were obtained from 13 patients (15 sites) undergoing surgical therapy. Tissues were homogenized at 4 degrees C on Tris saline buffer [1 g (5 ml)-1], homogenate was centrifuged and the resultant supernatants were used in assays. Antibody to collagen and Aa, Pg and Bf in tissue eluates and serum were determined by competitive enzyme linked immunosorbant assay (ELISA) and conventional ELISA respectively using an alkaline phosphatase/p-nitrophenyl phosphate enzyme-substrate system. Sera from age and sex matched healthy subjects and pooled human serum were used as controls. Antibody (Ab) levels in tissues and serum were standardized by concomitant albumin assay. Level of antibodies to collagen type 1 in tissue was significantly higher than in serum (P = 0.0001). Antibody levels in tissue to Pg were significantly higher than in serum (P = 0.0271). Ab levels to both Aa and Bf in tissues and serum were not significantly different from each other. These findings confirm the process of the local production of antibodies to autoantigen namely collagen type-1 and to bacterial antigens in the granulomatous tissues housed within the periodontal lesions in patients with periodontitis.

Locally produced anti-phosphorylcholine and anti-oxidized low-density lipoprotein antibodies in gingival crevicular fluid from aggressive periodontitis patients.

Sera from patients with periodontal attachment loss contain higher concentrations of IgG anti-phosphorylcholine (anti-PC) than sera from healthy subjects. Furthermore, a large proportion of plaque bacteria bear PC-containing surface antigens, implicating the oral flora as a source of immunogen for anti-PC. Additionally, anti-PC is cross-reactive with a variety of oral bacterial antigens and human antigens such as oxidized low-density lipoprotein (oxLDL). We hypothesized that, if the oral flora is a source of PC antigens, then we should be able to detect local anti-PC and anti-oxLDL production in gingival crevicular fluid (GCF). To test this, we collected 66 GCF samples from 15 patients with aggressive periodontitis and examined both the GCF samples and serum samples for their content of IgG anti-PC, IgG anti-LDL, and IgG anti-oxLDL by enzyme-linked immunosorbent assay. We also determined levels of anti-tetanus toxoid (anti-TT) as a non-oral antigen control. Serum and GCF concentrations of serum albumin (HSA) were also determined for use as a dilution marker. A conservative GCF:serum antibody ratio of greater than 1.5 was considered to be evidence of local antibody production. For the non-oral antigen TT, only one out of 62 samples contained locally produced antibody. Eight out of 64 samples (7 from a single subject) demonstrated local production of anti-LDL. In contrast, 28 out of 66 samples demonstrated local production of anti-PC, and 47 out of 66 samples contained locally produced anti-oxLDL. It was observed that A. actinomycetemcomitans strains containing or devoid of PC could absorb anti-oxLDL from human sera. Although there was a correlation between the ratios of anti-PC and anti-oxLDL (Spearman's rho = 0.35, P = 0.0037), local production of both antibodies was found in only 17 out of 65 samples, indicating that these antibodies are not always reflective of reactivity to the same antigens. The local production of anti-PC and anti-oxLDL further implicates the oral flora as a source of antigen that may mediate immune reactions of relevance to cardiovascular and other systemic diseases.

Immune deviation and ocular infections with varicella zoster virus

Since experimental, herpes simplex virus-induced acute retinal necrosis (ARN) develops in mice only if the mice fail to acquire virus-specific delayed hypersensitivity (DH) and despite their production of anti-viral antibodies (i.e. ACAID), I investigated whether a similar situation exists for patients with either varicella zoster virus (VZV)-induced ARN or anterior uveitis caused by VZV. Patients with either acute VZV-induced ARN, anterior uveitis with dermatitis (herpes zoster ophthalmicus, ZO-AU), or anterior uveitis without dermatitis (zoster sine herpete, ZSH-AU) were skin-tested with VZV to evaluate DH. The formal diagnoses of ARN associated with VZV, ZO-AU, and ZSH-AU were established by PCR analysis of the ocular samples and/or by the Goldmann-Witmer coefficient to determine levels of local antibody production. ARN, ZO-AU, and ZSH-AU activity were assessed clinically, and DH skin tests were repeated three months after onset when ocular recovery had taken place. All patients with VZV-induced skin disease alone (control group) displayed intense DH when tested with VZV antigen. In contrast, subsets of patients with ARN or ZO-AU displayed loss of VZV-specific DH. Patients with the most severe ARN or ZO-AU had the lowest DH responses to VZV antigens. Serum anti-VZV antibody titers were higher in ARN patients than in normal controls, and the anti-viral titer correlated inversely with the intensity of anti-VZV DH responses. VZV-specific DH responses were restored in patients who recovered from ARN. Patients with ZSH-AU also failed to display VZV-specific DH. The absence of DH reactivity to VZV antigens (i.e. immune deviation) appears to be a concomitant feature of VZV uveitis of high intensity, implying that virus-specific DH may interfere with the emergence of VZV-induced ARN or anterior uveitis.

Antibody-mediated protection against cytotoxic T-cell escape in coronavirus-induced demyelination.

C57BL/6 (B6) mice infected with mouse hepatitis virus (MHV) strain JHM develop a clinically evident, demyelinating encephalomyelitis. Infectious virus can be isolated from the spinal cords of these mice and is invariably mutated in the immunodominant CD8 T-cell epitope recognized in this strain. We showed previously that these persistently infected mice did not mount a measurable serum anti-MHV neutralizing antibody response. Here we show that cytotoxic T-lymphocyte (CTL) escape was not detected in MHV-infected BALB/b mice (H-2(b) haplotype), even though the same CD8 T-cell epitopes were recognized as in B6 mice. BALB/b mice had 25-fold more MHV-specific antibody-secreting cells in the central nervous system, the site of infection, than B6 mice, suggesting that local production of anti-MHV antibody contributed to this absence of CTL escape. Additionally, administration of anti-MHV neutralizing antibody to infected B6 mice suppressed the development of CTL escape mutants. These findings indicate a key role for the anti-MHV antibody response in suppressing virus replication, thereby minimizing the emergence and competitive advantage of CTL escape mutants.

Nonnecrotizing herpetic retinopathies masquerading as severe posterior uveitis

Objective Aqueous humor analysis can be performed in severe atypical forms of posterior uveitis unresponsive to conventional treatment to exclude a viral infection. Design Noncomparative interventional case series. Participants Thirty-seven immunocompetent patients seen with corticosteroid-resistant forms of posterior uveitis underwent extensive evaluation, including anterior chamber paracentesis, to rule out a nonnecrotizing viral retinopathy. Intervention Aqueous fluid samples were prospectively obtained. Polymerase chain reaction (PCR) and serologic evaluation of intraocular antibody production against herpesviruses were performed by molecular techniques and enzyme-linked immunosorbent assay. Main outcome measures Polymerase chain reaction and local antibody production for herpes simplex virus types 1 and 2, varicella-zoster virus, cytomegalovirus, and Epstein-Barr virus were determined on aqueous fluid samples. Results Viral infection was confirmed in 5 cases (13.5%). Clinical presentation included birdshot-like retinochoroidopathy, occlusive bilateral vasculitis, and cystoid macular edema. An antiviral regimen was initiated in all cases. Inflammation was stabilized, and steroid dosage could be significantly reduced. Conclusions Identification of a viral agent during severe posterior uveitis can dramatically change therapeutic management.

Review of Newcastle disease virus with particular references to immunity and vaccination

Newcastle disease (ND) is a highly contagious disease. The present paper deals with classification of ND virus (NDV), clinical signs and pathology, virus strain classification and molecular backgrounds for the pathogenicity. Major emphasis is reviewing immunity and vaccination. Clinical forms of the disease vary depending on many factors, but mainly on the virulence of Newcastle disease virus (NDV) strains. Virulent strains are considered List A pathogens by the ‘Office International des Epizooties' (OIE). The virulence has been traditionally determined using in vivo pathogenicity tests to distinguish between highly, moderately and low virulent isolates. More recently, molecular biological techniques like polymerase chain reaction and sequencing have been described to differentiate virulent from non-virulent strains.The systemic and mucosal immune systems are considered to function more or less independently. Systemic antibodies are essential elements in protection against ND, whereas the local antibodies limit multiplication of NDV at the site of entry. Cytotoxic T lymphocytes specific against NDV have been detected in the spleen of vaccinated birds, however, their contribution to protection remains to be elucidated. An increase of the number of various leukocyte subsets was noticed in the respiratory tract and the Harderian gland (HG), which favours involvement of local cellular immunity in the defence against NDV infection. It is tempting to speculate that the local lymphoid infiltrates are involved in first defence and that cytolytic cells clear virus by directly lysing infected target cells at the site of NDV inoculation. Secondly, various cell types, mainly T-lymphocytes and macrophages, may be equipped to produce a range of cytokines with antiviral activity and cytokines that stimulate B-lymphocytes to proliferate and differentiate into antibody-forming cells responsible for the local antibody production against NDV.Vaccination using inactivated virus induces mainly systemic immunity and usually protects birds from disease caused by virulent NDV strains. Mucosal vaccination would be more attractive, as it induces in addition to systemic immunity a local IgM, IgA, and IgG, response as well as a mucosal cellular immune response. These local responses contribute to protection against NDV infection of the respiratory tract mucosa, which is the first target tissue of infection with this virus.

Regional antibody and cellular immune responses to equine influenza virus infection, and particle mediated DNA vaccination

We have previously demonstrated that hemagglutinin (HA) gene vaccination and influenza virus infection generate protective antibody responses in equids. However, these antibody responses differ substantially in that particle mediated DNA vaccination does not induce an immunoglobulin A (IgA) response. A study was performed to investigate the regional immunoregulatory mechanisms associated with these different immune responses.Ponies were either vaccinated with equine HA DNA vaccines at skin and mucosal sites, infected with influenza virus or left untreated and influenza-specific antibody responses and protection from challenge infection was studied. In a subset of ponies, lymphocytes from peripheral blood (PBLs), nasopharyngeal mucosal tissue, or lymph nodes (LNLs) were collected for measurement of influenza virus-specific lymphoproliferative responses, local antibody production and IL-2, IL-4 and IFN-γ mRNA production by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).DNA vaccination and influenza virus infection induced humoral immunoglobulin Ga (IgGa) and immunoglobulin Gb (IgGb) production and lymphoproliferative responses that were positively correlated with IFN-γ mRNA production. However, there were marked differences in immune response in that only influenza infection induced an IgA response, and the regional distribution of lymphoproliferation, IFN-γ and antibody responses. Responses to DNA vaccination occurred in PBLs and in lymph nodes draining DNA vaccination sites, while influenza virus infection induced responses in PBLs and hilar LNLs. In summary, common features of immune responses to either influenza virus infection or DNA vaccination were virus-specific IgGa, IgGb and IFN-γ responses, which are associated with protection from infection, even when the regional distribution of these immune responses varied depending on the site of immune encounter.

Local immune responses to influenza virus infection in mice with a targeted disruption of perforin gene

The role of perforin in the local defense mechanisms against influenza virus infection was investigated. Mice deficient in the perforin gene (perforin −/−) were more susceptible to influenza virus infection than the ordinary wild-type C57BL/6 mice, showing an increased mortality with elevated virus growth and prolonged virus shedding. The lung parenchyma cells of perforin −/− mice showed no cytolytic activities of natural killer cells or virus-specific cytotoxic T lymphocytes in vitro, although the local antibody production system in the respiratory tract functioned well. In perforin −/− mice, the appearance of apoptotic degeneration in virally infected lung cells was delayed. This might cause cellular infiltration, including CD4, CD8, and CD19 positive cells, in the lung, peaking at day 8 after infection and maintaining a high level for a longer period. In the virus-induced local cytokine production, interferon-gamma (IFN-γ) was prominent. The adoptive transfer of immune-competent spleen cells from wild-type C57BL/6 mice achieved a complete protection of perforin −/− mice against lethal challenge infection. These results suggest that perforin plays a crucial role in the host defense system against influenza virus infection, especially in its early stage, by inducing apoptosis of virus-infected cells.

Testing ocular fluids in uveitis.

Infectious uveitis is a serious condition requiring rapid diagnosis and therapeutic management. Molecular tools, such as PCR and its variants, have significantly changed the diagnostic approach during the last 10 years. Presumed and empirical diagnosis should be excluded in the face of atypical clinical presentations, but also inadequate response to anti-inflammatory drugs. It is of particular importance to note that most of these tests need to be processed in specialized central laboratories to ensure the best sensitivity and specificity. Molecular techniques have their own limits, sometimes caused by inadequate primers or PCR-inhibitors and contaminants. PCR positivity means detection of pathogen DNA, but does not confirm a productive infection. Cultures from ocular fluids should be improved to obtain complementary data about pathogen life cycles and resistance to antibiotics or antivirals. Furthermore, diagnostic yield is significantly increased when PCR and local antibody production are associated, especially in viral infections. This step is of particular importance to identify new infectious entities in cases that are presently considered to be idiopathic or autoimmune disorders.

Chlamydia trachomatis in subfertile women undergoing uterine instrumentation: an alternative to direct microbial testing or prophylactic antibiotic treatment.

Chlamydia trachomatis is the major cause of tubal occlusion, and is also associated with IVF failure and spontaneous abortion. These infections are asymptomatic in most individuals and can persist in the genital tract for long periods of time in a form resistant to immune destruction. A significant percentage of couples seeking treatment for infertility might, therefore, harbour C. trachomatis in their genital tract. An unresolved question is what to do about this possible chlamydial persistence. Cervical, endometrial and semen samples can be tested for C. trachomatis and only positive individuals treated. Alternatively, all couples undergoing infertility treatment can receive prophylactic antibiotics. We advocate a third option, to screen and treat only individuals who are positive for systemic and/or local anti-chlamydial antibody production. Detection of species-specific C. trachomatis antibodies in peripheral blood will determine which individuals have been exposed to this organism and who, therefore, may be at risk for harbouring persistent forms. Identification of IgA antibodies in genital tract secretions may be an even better indicator of the presence of C. trachomatis in the genital tract. Circulating antibodies to the chlamydial 60kDa heat shock protein (hsp60) is a specific indicator of tubal occlusion and, furthermore, correlates with the continued presence of this micro-organism in the genital tract of non-human primates. Screening for both cervical IgA antibodies to C. trachomatis and serum IgG anti-chlamydial hsp60 appears to provide the best indication as to which women may be harbouring C. trachomatis.

Diagnosis of toxoplasmic retinochoroiditis with atypical clinical features

PURPOSE: To determine the value of aqueous humor analysis for confirming the diagnosis of ocular toxoplasmosis in patients who present with atypical clinical features and to relate the results of local antibody production and polymerase chain reaction (PCR) with the extent of active retinitis and the immune status of the patient. DESIGN: Retrospective case series. METHODS: Sixty-seven consecutive patients with retinitis or retinochoroiditis that was clinically consistent with atypical ocular toxoplasmosis underwent diagnostic anterior chamber paracentesis and serological studies. The aqueous humor was analyzed both by PCR to detect Toxoplasma gondii B1 gene and by the Goldman-Witmer coefficient to determine levels of local anti- T. gondii antibody production. RESULTS: In nine of the 67 cases, PCR was positive for T. gondii; seven of these were negative for local antibody production. All nine patients had illnesses associated with immunosuppression or advanced old age and all had active retinitis with a mean area of 11.5 disk areas (DA). Twenty-five of the remaining 58 cases were positive for local antibody production. These 25 had a mean area of active retinitis measuring 2.6 DA, and 24 of these patients were immunocompetent. All 34 cases with laboratory evidence of ocular toxoplasmosis diagnosed by either method responded to anti- T. gondii agents. The remaining 33 were negative for T. gondii infection by these two methods; some had laboratory evidence of other infections. CONCLUSIONS: Although in the present study, the sensitivity and specificity of the aqueous humor PCR and Goldman-Witmer coefficient could not be ascertained in the laboratory diagnosis of toxoplasmic retinochoroiditis, the PCR method appears to confirm the diagnosis in immunocompromised individuals with large atypical foci of retinitis. Conversely, determination of local antibody production may be appropriate for proper diagnosis in immunocompetent individuals presenting with small foci of retinitis.

Ocular toxoplasmosis antibodies in aqueous humor and serum

The diagnosis of ocular toxoplasmosis is mainly based on ophthalmological examination but might be difficult to establish in some cases. The purpose of our study was to evaluate the value of aqueous humor and serum analysis in ocular toxoplasmosis. We analyzed the avidity of toxoplasma-specific IgG in aqueous humor and serum samples from 50 patients with toxoplasmic retinochoroiditis, with 25 patients with uveitis posterior or panuveitis serving as controls. Specific intraocular antibody synthesis could be confirmed in 49 patients (98%). In two patients (8%) of the control group, antibody synthesis was detected (false positive). Forty-nine patients with diagnoses of ocular toxoplasmosis were positive for serum anti-T. gondii IgG, but only three patients had increased IgM levels. Analysis of local antibody production is a reliable method for confirming or excluding a suspected clinical diagnosis of toxoplasma retinochoroiditis. The determination of toxoplasma antibodies in the patients' serum is of limited value.

Aging impairs intestinal immunity

The elderly are characterized by immunosenescence accompanied by high rates of morbidity and mortality associated with infectious diseases. Despite suggestions that the mucosal immune compartment is relatively unaffected by aging, there are marked deficits in the intestinal mucosal immune responses of old animals and elderly humans. Little is known about the mechanism(s) whereby aging disrupts intestinal immunity. However, several events in the genesis of the intestinal immune response may be perturbed during aging. The first step is the uptake of antigens by specialized epithelial cells (M cells) that overlie the domes of Peyer's patches. We are unaware of any studies on the efficacy of antigen uptake in the intestine as a function of age. The effects of aging on the next step, antigen presentation by dendritic cells and lymphocyte isotype switching, have not been resolved. The third event is the maturation of immunoglobulin A (IgA) immunoblasts and their migration from the Peyer's patches to the intestinal mucosa. Quantitative immunohistochemical analyses suggest that the migration of these putative plasma cells to the intestinal effector site is compromised in old animals. Local antibody production by mature IgA plasma cells in the intestinal mucosa constitutes the fourth step. We recently reported that in vitro IgA antibody secretion by intestinal lamina propria lymphocytes from young and senescent rats is equivalent. The last event is the transport of IgA antibodies across the epithelial cells via receptor-mediated vesicular translocation onto the mucosal surface of the intestine. Receptor-binding assays did not detect age-associated declines in receptor number or binding affinity in either rodent or primate enterocytes as a function of donor age. Efforts to identify the mechanism(s) responsible for the age-related decline in intestinal mucosal immune responsiveness may benefit by focusing on the homing of IgA immunoblasts to the effector site.

Expression of recombination-activating genes and terminal deoxynucleotidyl transferase and secondary rearrangement of immunoglobulin kappa light chains in rheumatoid arthritis synovial tissue.

Lymphocytic infiltrates in rheumatoid arthritis (RA) synovium often resemble lymphoid follicles and contain clonally related Ig transcripts, suggesting in situ antigen-dependent B cell selection. Recent reports have shown expression of recombination-activating genes (RAGs) and concurrent secondary rearrangement of Ig genes in normal peripheral lymphoid organs (receptor revision). We sought to determine if RAG-mediated receptor revision of Ig kappa light chains occurs in B cells within the RA synovium. Because we previously reported enhanced N-region addition at V(L)-J(L) joins in clonally expanded light-chain transcripts from RA synovium, we also sought expression of terminal deoxynucleotidyl transferase (TdT), which is normally expressed only in B cell precursors or immature B cells. Reverse transcription-polymerase chain reaction (PCR) was used to detect RAG and TdT transcripts from unselected and B cell-enriched synovial and peripheral blood mononuclear cells obtained from 12 RA patients. Activity of RAG protein was sought using ligation-mediated PCR to detect recombination intermediates, and immunohistochemistry was performed to identify RAG+ cells within synovia. We found evidence of RAG-mediated secondary Ig kappa light chain rearrangements in about one-third of RA synovia. TdT expression was found in several samples, but did not correlate with RAG expression. RAG-mediated secondary Ig rearrangements of kappa light chains may contribute to the local production of antibodies to autoantigens (e.g., rheumatoid factor) or exogenous antigens, or it may represent a failed attempt at immune tolerance. TdT expression suggests the presence of immature B cells in RA synovia. These findings have important implications for the local generation of antibodies in RA and other chronic inflammatory diseases.

Anterior uveitis with sectoral iris atrophy in the absence of keratitis

Objective To determine the cause and describe the clinical features of unilateral anterior uveitis with sectoral atrophy of the iris in the absence of associated keratitis. Design Retrospective, observational case series. Participants Thirty-one patients with unilateral anterior uveitis with sectoral iris atrophy and without (previous) keratitis. Methods The patients were selected from our database of 592 patients with anterior uveitis. Main outcome measures We reviewed the clinical data on the 31 patients and the results of diagnostic anterior chamber fluid analysis for 24 of the 31 patients. Specifically, production of local antibodies against herpes simplex virus (HSV) and varicella zoster virus (VZV) was determined and the polymerase chain reaction was performed to demonstrate the DNA of HSV, VZV, and cytomegalovirus (CMV) in the aqueous samples. Results Main clinical characteristics of anterior uveitis with iris atrophy included unilateral involvement with a prolonged course and recurrent exacerbations in all cases. Elevated intraocular pressure during intraocular inflammation occurred in 90% of patients (28 of 31). Visual outcome was favorable because 29 of 31 patients (94%) retained a visual acuity of 20/32 or more. The causal agent was identified as HSV in 83% (20 of 24) and VZV in 13% (3 of 24) and was inconclusive in one case. The patients with HSV uveitis were younger than those with VZV uveitis (mean age at onset 34 and 65 years, respectively; P = 0.0056). Conclusions Unilateral anterior uveitis with sectoral atrophy of the iris without associated (previous) keratitis is a distinct entity among herpetic eye diseases. Recurrent unilateral anterior uveitis with iris atrophy and/or elevated intraocular pressure has most likely been caused by HSV.

Lack of association between human immunodeficiency virus type 1 antibody in cervicovaginal lavage fluid and plasma and perinatal transmission, in Thailand.

To determine the association between human immunodeficiency virus type 1 (HIV)-specific antibody and RNA levels in cervicovaginal lavage (CVL) samples and plasma, zidovudine treatment, and perinatal transmission, HIV subtype E gp160-specific IgG and IgA were serially measured in a subset of 74 HIV-infected women in a placebo-controlled trial of zidovudine, beginning at 36 weeks of gestation. HIV IgG was detected in 100% of plasma and 97% of CVL samples; HIV IgA was consistently detected in 62% of plasma and 31% of CVL samples. Antibody titers in CVL samples correlated better with the RNA level in CVL samples than with plasma antibody titers. Zidovudine did not affect antibody titers. Perinatal HIV transmission was not associated with antibody in CVL samples or plasma. HIV-specific antibody is present in the cervicovaginal canal of HIV-infected pregnant women; its correlation with the RNA level in CVL fluid suggests local antibody production. However, there was no evidence that these antibodies protected against perinatal HIV transmission.