Lacrimal Cytokines Assessment in Subjects Exposed to Different Levels of Ambient Air Pollution in a Large Metropolitan Area.

BACKGROUND: Air pollution is one of the most environmental health concerns in the world and has serious impact on human health, particularly in the mucous membranes of the respiratory tract and eyes. However, ocular hazardous effects to air pollutants are scarcely found in the literature. DESIGN: Panel study to evaluate the effect of different levels of ambient air pollution on lacrimal film cytokine levels of outdoor workers from a large metropolitan area. METHODS: Thirty healthy male workers, among them nineteen professionals who work on streets (taxi drivers and traffic controllers, high pollutants exposure, Group 1) and eleven workers of a Forest Institute (Group 2, lower pollutants exposure compared to group 1) were evaluated twice, 15 days apart. Exposure to ambient PM2.5 (particulate matter equal or smaller than 2.5 µm) was 24 hour individually collected and the collection of tears was performed to measure interleukins (IL) 2, 4, 5 and 10 and interferon gamma (IFN-γ) levels. Data from both groups were compared using Student's t test or Mann- Whitney test for cytokines. Individual PM2.5 levels were categorized in tertiles (lower, middle and upper) and compared using one-way ANOVA. Relationship between PM2.5 and cytokine levels was evaluated using generalized estimating equations (GEE). RESULTS: PM2.5 levels in the three categories differed significantly (lower: ≤22 µg/m3; middle: 23-37.5 µg/m3; upper: >37.5 µg/m3; p<0.001). The subjects from the two groups were distributed unevenly in the lower category (Group 1 = 8%; Group 2 = 92%), the middle category (Group 1 = 89%; Group 2 = 11%) and the upper category (Group 1 = 100%). A significant relationship was found between IL-5 and IL-10 and PM2.5 levels of the group 1, with an average decrease of 1.65 pg/mL of IL-5 level and of 0.78 pg/mL of IL-10 level in tear samples for each increment of 50 µg/m3 of PM2.5 (p = 0.01 and p = 0.003, respectively). CONCLUSION: High levels of PM2.5 exposure is associated with decrease of IL-5 and IL-10 levels suggesting a possible modulatory action of ambient air pollution on ocular surface immune response.

Expression of IL-8, IL-6 and IL-1ß in tears as a main characteristic of the immune response in human microbial keratitis.

Corneal infections are frequent and potentially vision-threatening diseases, and despite the significance of the immunological response in animal models of microbial keratitis (MK), it remains unclear in humans. The aim of this study was to describe the cytokine profile of tears in patients with MK. Characteristics of ocular lesions such as size of the epithelial defect, stromal infiltration, and hypopyon were analyzed. Immunological evaluation included determination of interleukine (IL)-1ß, IL-6, IL-8, IL-10, IL-12 and tumor necrosis factor (TNF)-α in tear samples obtained from infected eyes of 28 patients with MK and compared with their contralateral non-infected eyes. Additionally, frequency of CD4+, CD8+, CD19+ and CD3-CD56+ cells was also determined in peripheral blood mononuclear cells in patients with MK, and compared with 48 healthy controls. Non-significant differences were observed in the size of the epithelial defect, stromal infiltration, and hypopyon. Nevertheless, we found an immunological profile apparently related to MK etiology. IL-8 > IL-6 in patients with bacterial keratitis; IL-8 > IL-6 > IL-1ß and increased frequency of circulating CD3-CD56+ NK cells in patients with gram-negative keratitis; and IL-8 = IL-6 > IL-1ß in patients with fungal keratitis. Characterization of tear cytokines from patients with MK could aid our understanding of the immune pathophysiological mechanisms underlying corneal damage in humans.

The ocular surface: from physiology to the ocular allergic diseases.

Allergic conjunctivitis (AC) is an inflammation of the conjunctiva secondary to an immune response to exogenous antigens, usually called allergens. In fact, AC is a syndrome that involves the entire ocular surface, including conjunctiva, lids, cornea, and tear film. The signs and symptoms of AC have a meaningful effect on comfort and patient health, and could be influenced by environment, genetics and immune regulation mechanisms, all of which work together in a complex immunological homeostasis. Dysregulation in such immune responses could turn into a variety of ocular allergic diseases (OAD). This review describes some of the current understanding of cellular and molecular pathways involved in different OAD.

La conjuntivitis alérgica es la inflamación de la conjuntiva secundaria a una respuesta inmunitaria contra antígenos exógenos, usualmente llamados alergenos. De hecho, la conjuntivitis alérgica es un síndrome que involucra la totalidad de la superficie ocular, incluyendo la conjuntiva, los párpados, la córnea y la película lagrimal. Los signos y síntomas de la conjuntivitis alérgica tienen un efecto significativo en el bienestar y salud del paciente y pueden ser influidos por el ambiente, la genética y mecanismos de regulación inmunológicos, todos los cuales trabajan en conjunto en una compleja homeostasia inmunológica. La disregulación de estos mecanismos puede desembocar en una gran variedad de enfermedades alérgicas oculares. Esta revisión describe algunos de los conocimientos celulares y moleculares actuales, involucrados en las diferentes enfermedades alérgicas oculares.

Evaluation of anti-Moraxella bovis pili immunoglobulin-A in tears following intranasal vaccination of cattle.

Infectious bovine keratoconjunctivitis (IBK) is a highly contagious ocular disease of cattle caused by Moraxella bovis (Mb). Parenterally administered immunogens used to prevent the disease do not offer complete protection possibly because they stimulate a poor ocular mucosal secretory response, in which locally secreted immunoglobulin-A (sIgA) is one of the main components. The principal aim of this study was to evaluate by an indirect enzyme linked immunosorbent assay (ELISA), the local ocular mucosal sIgA response against Mb purified pili, produced after intranasal inoculation of experimental vaccines. Pili were adjuvanted by several different adjuvants (QuilA, Marcol Arlacel, Marcol Span, microencapsulated pili with PLGA polymers). Results were compared to sIgA response produced by adjuvant placebo inoculations and by IBK natural infection. Significantly higher anti-pili IgA response (p<0.05) was detected in calves vaccinated intranasally with pili QuilA and pili Marcol Span compared to control calves, although this specific immune response did not seem to be related to protection against Mb infection or typical IBK lesion development.

Ocular toxoplasmosis: evaluation of lacrimal-specific secretory IgA levels in both patients with active and inactive phases of the disease.

Ocular toxoplasmosis can result in recurrent uveitis. Studies have shown that a correlation between active ocular toxoplasmosis and the presence of anti-Toxoplasma gondii secretory IgA (SIgA) in tears. This study compares anti-T. gondii SIgA levels in patients' tears during the acute and inactive phases of toxoplasmic uveitis. Twenty-nine positive tear specific SIgA for T. gondii patients with acute toxoplasmic uveitis were selected and were followed-up for at least two years, when the anti-T. gondii SIgA tears levels were determined. Specific SIgA for T. gondii was negative in 22 patients (75.86%) and positive in seven patients (24.13%) of whom six (85.7%) were followed over three years. Average SIgA levels during the acute phase are 1.54 and decrease significantly to 0.72 (p = 0.0001) during the inactive phase of disease. Because anti-T. gondii SIgA in the tear is negative in 75.86% of patients after the acute phase of infection, T. gondii SIgA levels may be used as a complementary diagnostic marker for active ocular toxoplasmosis.

Ocular toxoplasmosis: evaluation of lacrimal - specific secretory IgA levels in both patients with active and inactive phases of the disease

Ocular toxoplasmosis can result in recurrent uveitis. Studies have shown that a correlation between active ocular toxoplasmosis and the presence of anti-Toxoplasma gondii secretory IgA (SIgA) in tears. This study compares anti-T. gondii SIgA levels in patients' tears during the acute and inactive phases of toxoplasmic uveitis. Twenty-nine positive tear specific SIgA for T. gondii patients with acute toxoplasmic uveitis were selected and were followed-up for at least two years, when the anti-T. gondii SIgA tears levels were determined. Specific SIgA for T. gondii was negative in 22 patients (75.86 percent) and positive in seven patients (24.13 percent) of whom six (85.7 percent) were followed over three years. Average SIgA levels during the acute phase are 1.54 and decrease significantly to 0.72 (p = 0.0001) during the inactive phase of disease. Because anti-T. gondii SIgA in the tear is negative in 75.86 percent of patients after the acute phase of infection, T. gondii SIgA levels may be used as a complementary diagnostic marker for active ocular toxoplasmosis.

Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA.

Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA) antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with ocular toxoplasmosis. Tears from 156 patients with active posterior uveitis were analysed; 82 of them presented characteristics of ocular toxoplasmosis (standard lesion) and 74 patients presented uveitis due to other aetiologies. Cases of active posterior uveitis were considered standard when a new inflammatory focus satellite to old retinochoroidal scars was observed. The determination of anti-T. gondii sIgA was made using an ELISA test with crude tachyzoite antigenic extracts. Tears were collected without previous stimulation. Detection of sIgA showed 65.9% sensitivity (95% CI = 54.5-74.4), 71.6% specificity (95% CI = 59.8-81.2), a positive predictive value of 72% (95% CI = 60.3-81.5) and a negative predictive value of 65.4% (95% CI = 54.0-75.4). sIgA reactivity was higher in the tears of patients with active posterior uveitis due to T. gondii (p < 0.05). The test is useful for differentiating active posterior uveitis due to toxoplasmosis from uveitis caused by other diseases.

Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA

Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA) antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with ocular toxoplasmosis. Tears from 156 patients with active posterior uveitis were analysed; 82 of them presented characteristics of ocular toxoplasmosis (standard lesion) and 74 patients presented uveitis due to other aetiologies. Cases of active posterior uveitis were considered standard when a new inflammatory focus satellite to old retinochoroidal scars was observed. The determination of anti-T. gondii sIgA was made using an ELISA test with crude tachyzoite antigenic extracts. Tears were collected without previous stimulation. Detection of sIgA showed 65.9 percent sensitivity (95 percent CI = 54.5-74.4), 71.6 percent specificity (95 percent CI = 59.8-81.2), a positive predictive value of 72 percent (95 percent CI = 60.3-81.5) and a negative predictive value of 65.4 percent (95 percent CI = 54.0-75.4). sIgA reactivity was higher in the tears of patients with active posterior uveitis due to T. gondii (p < 0.05). The test is useful for differentiating active posterior uveitis due to toxoplasmosis from uveitis caused by other diseases.

Immunogenicity of recombinant Omp31 from Brucella melitensis in rams and serum bactericidal activity against B. ovis.

Detergent-extracted recombinant Omp31 (rOmp31 extract) from Brucella melitensis produced in Escherichia coli was previously identified as a protective immunogen against B. ovis in mice. In this study, we evaluated the immunogenicity of rOmp31extract in rams. This immunogen was emulsified in an oil adjuvant and administered three times with 4 and 8 weeks intervals. Antibody response was measured in serum by whole B. ovis ELISA. Specific antibodies to purified rOmp31 (pET-Omp31) were detected by Western blotting and indirect ELISA. In addition, isotype specific antibodies were measured in tears. Serum bactericidal activity against B. ovis in the presence of complement was measured in vitro. Cellular immune response was explored by intradermal testing with purified rOmp31. Immunization with rOmp31 extract induced IgG specific antibodies in serum able to bind to whole B. ovis cells. Furthermore, strong inhibition in a competitive ELISA (with an Omp31-specific monoclonal antibody) suggested that a proportion of Omp31-specific antibodies were directed against a loop containing a protective epitope. Serum antibodies killed efficiently B. ovis in vitro in the presence of either guinea pig or ovine serum. Tears had both IgG and IgA antibodies to equivalent titers. Finally, immunized rams showed skin reactivity to Omp31. These data demonstrate that B. melitensis Omp31, a protective antigen identified in the mouse model, induces antibody and cellular immune mechanisms in sheep.

Lacrimal secretory IgA in active posterior uveitis induced by Toxoplasma gondii.

It is quite difficult to diagnose active toxoplasmosis in patients with ocular toxoplasmosis. Active posterior uveitis presumably due to Toxoplasma gondii infection (APUPT) is seldom produced during a prime-infection; hence most patients do not show high IgM antibodies. High levels of IgA have been described in active toxoplasmosis. The purpose of this study was to investigate possible association between APUPT and the specific anti-parasite sIgA in tears. The study was carried out as case-control. Tears of 25 clinically confirmed APUPT patients and 50 healthy control subjects were analyzed. All were IgG seropositive. Specific sIgA was determined by ELISA assay using T. gondii RH strain crude extract. Anti-T. gondii sIgA was found in 84% of the cases and in 22% of the control subjects. The intensity of the reaction was higher in APUPT cases (P = 0.007). There was strong association between APUPT patients and lacrimal sIgA (odds-ratio 18.61, P = 0.0001). ELISA test sensitivity was 84% and specificity 78%. Our data suggest that anti-T.gondii secretory IgA found in tears may become an important marker for active ocular toxoplasmosis.

Infectious bronchitis: effect of viral doses and routes on specific lacrimal and serum antibody responses in chickens.

An enzyme-linked immunosorbent assay was used to measure the effect of various infectious bronchitis virus (IBV) (strain H-120) vaccine doses and routes of immunization on specific lacrimal and serum antibody responses. The results of the first trial showed that the maximum dose, 10(6) median embryo infective doses (EID50s), delivered by the ocular route elicited both a systemic and a local antibody response in the vaccinated chickens. Lower doses of vaccinal virus, 10(4) (median dose) and 10(2) (minimum dose) EID50 delivered by the same route did not induce a detectable systemic antibody response. A significant increase of IBV-specific lacrimal IgA was elicited by both the maximum and the median vaccine doses. The low vaccine dose (10(2) EID50) did not induce a detectable increase of lacrimal IgA. In a second trial approximately the same vaccine dose was administered to different chicken groups by ocular instillation, drinking water, spray, and cloaca. The results showed that all routes of vaccination tested, including the cloacal route, resulted in an increase of specific serum antibodies. Higher IgG levels were detected throughout the experimental period after vaccination by the ocular route as compared with vaccination via the drinking water. All routes of vaccination tested resulted in an increase of specific IgA in lacrimal fluid. The vaccine application methods spray, ocular instillation, and drinking water induced similar lacrimal IgA responses.

A chlamydial major outer membrane protein extract as a trachoma vaccine candidate.

PURPOSE: As shown in infected humans and in animal models of chlamydial infection, the major outer membrane protein (MOMP) of Chlamydia trachomatis is immunogenically potent. The purpose of this investigation was to test in the cynomolgus monkey model of trachoma a new extract of MOMP as a candidate vaccine against ocular chlamydial infection. METHOD: The nonionic detergent octyl-beta-D glucopyranoside (OGP) was used to extract MOMP from purified C. trachomatis (serovar C) elementary bodies. Protective immunization with OGP-MOMP by mucosal and systemic routes was compared in the cynomolgus monkey model of trachoma. All control and immunized monkeys were challenged by topical application of infectious C. trachomatis to the conjunctivae 35 days after the initiation of immunization. RESULTS: Immunization with OGP-extracted MOMP successfully induced chlamydia-specific local and systemic immunity to MOMP and to whole organism before challenge and early clearance of infection by systemically immunized monkeys. Although ocular disease was not significantly reduced in either immunized group compared to control animals, the lowest clinical and microbiologic disease scores developed in two animals in the mucosal group with the highest immunoglobulin A tear antibody titers at day 0 to 14, whereas higher tear and serum immunoglobulin G correlated with reduced disease in the systemically immunized group. CONCLUSIONS: These data demonstrate that despite evidence of vigorous MOMP-specific and other chlamydia-specific serologic and cell-mediated immunity, as well as anamnestic serologic responses to chlamydia, vaccination with OGP-MOMP was only partially protective against chlamydial ocular disease. The partial protection correlated best with tear immunoglobulin A responses after mucosal immunization and with local and systemic immunoglobulin G responses after peripheral immunization, suggesting that alternative chlamydial antigens may have to be considered in future vaccine development to induce more effective protective immunity and that evaluation of efficacy must be appropriate to route of immunization.

Esclerosis múltiple test de los 4 humores: líquido cefalorraquideo, suero, lágrimas y saliva / Multiple sclerosis test or the 4 humors: cerebrospinal fluid serum, tears and saliva

Se estudiaron 4 fluidos biológicos de fácil acceso a la exploración inmune (líquido cefalorraquideo, suero, lágrimas y saliva) en 25 pacientes con Esclerosis Multiple (EM) obtenidos durante un empuje clínico de la enfermedad. El nivel del receptor de Interleukina-2 soluble (RsIL-2) se encontró significativamente aumentado en por lo menos 3 de estos 4 fluidos, en comparación con el de los controles normales. La sensibilidad y especificidad de su determinación para el diagnóstico de la afección, fue mayor que la de otros parámetros inmunoquímicos -distribución oligoclonal (DO) de inmunoglobulinas (Ig), disbalance de cadenas livianas- y estudios electrofisiológicos -potenciales evocados-. Este método es utilizado para establecer un diagnóstico más certero de la Esclerosis Múltiple así como también para monitorizar su actividad biológica junto con la resonancia magnética nuclear (RMN)(AU)

Esclerosis multiple test de los 4 humores: liquido cefalorraquideo, suero, lagrimas y saliva / Multiple sclerosis test or the 4 humors: cerebrospinal fluid serum, tears and saliva

Se estudiaron 4 fluidos biologicos de facil acceso a la exploracion inmune (liquido cefalorraquideo, suero, lagrimas y saliva) en 25 pacientes con Esclerosis Multiple (EM) obtenidos durante un empuje clinico de la enfermedad. El nivel del receptor de Interleukina-2 soluble (RsIL-2) se encontro significativamente aumentado en por lo menos 3 de estos 4 fluidos, en comparacion con el de los controles normales. La sensibilidad y especificidad de su determinacion para el diagnostico de la afeccion, fue mayor que la de otros parametros inmunoquimicos -distribucion oligoclonal (DO) de inmunoglobulinas (Ig), disbalance de cadenas livianas- y estudios electrofisiologicos -potenciales evocados-. Este metodo es utilizado para establecer un diagnostico mas certero de la Esclerosis Multiple asi como tambien para monitorizar su actividad biologica junto con la resonancia magnetica nuclear (RMN)

The clinical performance of the Surevue, Focus, and CSI daily wear lenses in allergic and normal contact lens patients.

Sixty-three daily wear contact lens patients (42 non-allergic [66.7%] and 21 allergic [33.3%]) were randomized into three groups (CSI vs. Surevue; Focus vs. CSI; Focus vs. Surevue) and evaluated every two weeks for six months with regard to patient preferences and ranked symptom responses for 14 variables, including overall contact lens preference. Allergic patients (43.9 +/- 94.3 ng/mL) revealed significantly higher (P = .011) tear IgE levels versus non-allergic patients (15.6 +/- 25.8 ng/mL). Non-allergic patients preferred CSI over Surevue for 12 of 14 variables including overall lens preference (P < .001); CSI over Focus for 6 of 14 variables, including overall lens preference (P < .009); and Focus over Surevue for 13 of 14 variables, including overall lens preference (P < .001). In contrast, allergic patients preferred Focus over CSI for 8 of 14 variables; CSI over Surevue for 4 of 14 variables, including overall lens preference (P < .001); Focus over Surevue for 4 of 14 variables, including overall lens preference (P < .001). Surevue and Focus lenses were replaced at 4 and 8 week intervals, respectively. No significant differences in the ranked symptom responses were observed for Surevue at 4 weeks versus 2 weeks, or for Focus at 8 weeks versus 4 weeks. A regression of time versus symptom responses for non-allergic patients wearing the CSI lens revealed decreasing symptoms over time, while the allergic patients showed no significant difference.(ABSTRACT TRUNCATED AT 250 WORDS)

Dosagem de lisozima na lágrima de recém-nascidos / Lysozyme concentration in tears of newborn babies

A concentraçäo de lisozima lacrimal foi medida pelo método da lisoplaca em 47 recém-nascidos divididos em 3 grupos; 22 de termo, 11 de termo com baixo peso e 14 prematuros. A concentraçäo de lisozima foi maior no grupo de termo comparando-se com o grupo de baixo peso e prematuros e seus valores aumentaram com peso e idade gestacional dos recém-nascidos

Estudo imunológico da lágrima nas granulomatoses de causa inalatória / Immunological study of lacrimal secretion in inhalatory induced granulomatosis

O segmento anterior do globo ocular, mais particularmente a conjuntiva, constitui um modelo experimental de "órgäo de choque" em relaçäo a agentes agressores ambienciais. Face à hipótese de existir uma reaçäo imunológica de defesa, os autores estudam em particular a vertente humoral desta. Neste sentido, analisam um grupo de doentes com exposiçäo aerogénea, alguns deles com manifestaçöes patológicas, classificadas por outros como Granulomatoses de Causa Inalatória. Säo doseados, na secreçäo lacrimal, as imunoglobulinas dos tipos A, G, E e M. Comentam-se os resultados