Hyaluronic acid biomaterial for human tissue-engineered skin substitutes: Preclinical comparative in vivo study of wound healing.

BACKGROUND: There is not an ideal biomaterial for tissue-engineered skin substitutes (TESSs), and most of the studies or existing therapies use xenogeneic origin natural biomaterials or biosynthetic scaffolds. OBJECTIVE: To analyse clinical, histological integration and homeostasis parameters of a human TESS manufactured with fibrin-hyaluronic acid biomaterial (HA-Skin), grafted in immunodeficient mice for 8 weeks, and compared with the gold standard treatment (Autograft), a human TESS manufactured with fibrin-agarose biomaterial (AG-Skin) and secondary wound healing dressings. METHODS: Human TESSs and autografts were implanted into BALB/c mice after surgical excision. Secondary wound healing approach was achieved with biosynthetic collagen wound dressing (Biobrane® ) and fibrin-hyaluronic acid or fibrin-agarose biomaterial without cells (Total N = 44). Clinical integration and homeostasis parameters were evaluated every two weeks for two months. Histological and immunohistochemical analyses were performed four and eight weeks after grafting. RESULTS: HA-Skin, AG-Skin and Autograft groups showed a proper clinical integration and epithelization eight weeks later. Scar evaluation revealed better results for Autograft and HA-Skin. Homeostasis analysis indicated similar values of transepidermal water loss and elasticity between HA-Skin (6.42 ± 0.75 g/h/m2 , 0.42 ± 0.08 AU), Autograft (6.91 ± 1.28 g/h/m2 , 0.40 ± 0.08 AU) and healthy mouse skin (6.40 ± 0.43 g/h/m2 , 0.35 ± 0.03 AU). Histological results showed that human TESSs and autografts presented better skin structuration and higher expression of cytokeratins. CONCLUSIONS: This study suggests that human TESS based on fibrin-hyaluronic acid biomaterial could be suitable for clinical application in the treatment of several dermatological pathologies (wound healing).

Uso da artéria torácica interna para tratamento cirúrgico de origem anômalo da artéria coronária esquerda: relato de caso / Use of the internal thoracic artery for surgical treatment of anomalous origin of the left coronary artery: case report

A origem anômala da artéria coronária esquerda do tronco pulmonar (Síndrome de Bland-White-Garland ou "Anomalous origin of Coronary Artery" (ALCAPA)) é uma anomalia congênita rara que afeta 1/300.000 nascidos vivos. Em 5% dos casos pode ser acompanhada com outra malformação congênita. Apresenta mortalidade de até 85% no primeiro ano de vida. A sintomatologia se relaciona a isquemia miocárdica e insuficiência cardíaca, sendo a correção cirúrgica o tratamento curativo. As principais técnicas cirurgicas são: reimplante do óstio coronário na aorta, interposição de artéria subclávia esquerda (Cirurgia Meyer), neo-tunelização (cirurgia de Takeuchi) e "bypass" coronário. Apresentamos um caso de paciente avaliado aos 9 meses de idade e 12 kg com quadro clínico de cianose durante as mamadas e dispnéia aos moderados esforços, sem outros antecedentes. Ao exame físico foi identificado sopro sistólico em foco pulmonar e aórtico. Realizou ecocardiograma e angiografia coronária sendo diagnosticado origem anômalo da artéria coronária esquerda. Sendo portanto submetido tratamento cirúrgico através da técnica "bypass" coronário utilizando artéria torácica interna esquerda. O paciente realiza seguimento anual no serviço há 21 anos apresentando evolução favorável. (AU)

MEPROCS framework for Craniofacial Superimposition: Validation study.

Craniofacial Superimposition (CFS) involves the process of overlaying a skull with a number of ante-mortem images of an individual and the analysis of their morphological correspondence. The lack of unified working protocols and the absence of commonly accepted standards, led to contradictory consensus regarding its reliability. One of the more important aims of 'New Methodologies and Protocols of Forensic Identification by Craniofacial Superimposition (MEPROCS)' project was to propose a common framework for CFS, what can be considered the first international standard in the field. The framework aimed to serve as a roadmap for avoiding particular assumptions that could bias the process. At the same time, it provides some empirical support to certain practices, technological means, and morphological criteria expected to facilitate the application of the CFS task and to improve its reliability. In order to confirm the utility and potential benefits of the framework use, there is a need to empirically evaluate it in CFS identification scenarios as close as possible to the reality. Thus, the purpose of this study is to validate the CFS framework developed. For that aim 12 participants were asked to report about a variable number of CFS following all the recommendations of the framework. The results are analysed and discussed according to the framework understanding and fulfilment, the participants' performance, and the correlation between expected decisions and those given by the participants. In view of the quantitative results and qualitative examination criteria we can conclude that those who follow the MEPROCS recommendations improve their performance.

Study on the performance of different craniofacial superimposition approaches (II): Best practices proposal.

Craniofacial superimposition, although existing for one century, is still a controversial technique within the scientific community. Objective and unbiased validation studies over a significant number of cases are required to establish a more solid picture on the reliability. However, there is lack of protocols and standards in the application of the technique leading to contradictory information concerning reliability. Instead of following a uniform methodology, every expert tends to apply his own approach to the problem, based on the available technology and deep knowledge on human craniofacial anatomy, soft tissues, and their relationships. The aim of this study was to assess the reliability of different craniofacial superimposition methodologies and the corresponding technical approaches to this type of identification. With all the data generated, some of the most representative experts in craniofacial identification joined in a discussion intended to identify and agree on the most important issues that have to be considered to properly employ the craniofacial superimposition technique. As a consequence, the consortium has produced the current manuscript, which can be considered the first standard in the field; including good and bad practices, sources of error and uncertainties, technological requirements and desirable features, and finally a common scale for the craniofacial matching evaluation. Such a document is intended to be part of a more complete framework for craniofacial superimposition, to be developed during the FP7-founded project MEPROCS, which will favour and standardize its proper application.

Study on the performance of different craniofacial superimposition approaches (I).

As part of the scientific tasks coordinated throughout The 'New Methodologies and Protocols of Forensic Identification by Craniofacial Superimposition (MEPROCS)' project, the current study aims to analyse the performance of a diverse set of CFS methodologies and the corresponding technical approaches when dealing with a common dataset of real-world cases. Thus, a multiple-lab study on craniofacial superimposition has been carried out for the first time. In particular, 26 participants from 17 different institutions in 13 countries were asked to deal with 14 identification scenarios, some of them involving the comparison of multiple candidates and unknown skulls. In total, 60 craniofacial superimposition problems divided in two set of females and males. Each participant follow her/his own methodology and employed her/his particular technological means. For each single case they were asked to report the final identification decision (either positive or negative) along with the rationale supporting the decision and at least one image illustrating the overlay/superimposition outcome. This study is expected to provide important insights to better understand the most convenient characteristics of every method included in this study.

Ground truth data generation for skull-face overlay.

Objective and unbiased validation studies over a significant number of cases are required to get a more solid picture on craniofacial superimposition reliability. It will not be possible to compare the performance of existing and upcoming methods for craniofacial superimposition without a common forensic database available for the research community. Skull-face overlay is a key task within craniofacial superimposition that has a direct influence on the subsequent task devoted to evaluate the skull-face relationships. In this work, we present the procedure to create for the first time such a dataset. We have also created a database with 19 skull-face overlay cases for which we are trying to overcome legal issues that allow us to make it public. The quantitative analysis made in the segmentation and registration stages, together with the visual assessment of the 19 face-to-face overlays, allows us to conclude that the results can be considered as a gold standard. With such a ground truth dataset, a new horizon is opened for the development of new automatic methods whose performance could be now objectively measured and compared against previous and future proposals. Additionally, other uses are expected to be explored to better understand the visual evaluation process of craniofacial relationships in craniofacial identification. It could be very useful also as a starting point for further studies on the prediction of the resulting facial morphology after corrective or reconstructive interventionism in maxillofacial surgery.

Dispersion assessment in the location of facial landmarks on photographs.

The morphological assessment of facial features using photographs has played an important role in forensic anthropology. The analysis of anthropometric landmarks for determining facial dimensions and angles has been considered in diverse forensic areas. Hence, the quantification of the error associated to the location of facial landmarks seems to be necessary when photographs become a key element of the forensic procedure. In this work, we statistically evaluate the inter- and intra-observer dispersions related to the facial landmark identification on photographs. In the inter-observer experiment, a set of 18 facial landmarks was provided to 39 operators. They were requested to mark only those that they could precisely place on 10 photographs with different poses (frontal, oblique, and lateral views). The frequency of landmark location was studied together with their dispersion. Regarding the intra-observer evaluation, three participants identified 13 facial points on five photographs classified in the frontal and oblique views. Each landmark location was repeated five times at intervals of at least 24 h. The frequency results reveal that glabella, nasion, subnasale, labiale superius, and pogonion obtained the highest location frequency in the three image categories. On the contrary, the lowest rate corresponds to labiale inferius and menton. Meanwhile, zygia, gonia, and gnathion were significantly more difficult to locate than other facial landmarks. They produced a significant effect on the dispersion depending on the pose of the image where they were placed, regardless of the type of observer that positioned them. In particular, zygia and gonia presented a statistically greater variation in the three image poses, while the location of gnathion is less precise in oblique view photographs. Hence, our findings suggest that the latter landmarks tend to be highly variable when determining their exact position.

Computer vision and soft computing for automatic skull-face overlay in craniofacial superimposition.

Craniofacial superimposition can provide evidence to support that some human skeletal remains belong or not to a missing person. It involves the process of overlaying a skull with a number of ante mortem images of an individual and the analysis of their morphological correspondence. Within the craniofacial superimposition process, the skull-face overlay stage just focuses on achieving the best possible overlay of the skull and a single ante mortem image of the suspect. Although craniofacial superimposition has been in use for over a century, skull-face overlay is still applied by means of a trial-and-error approach without an automatic method. Practitioners finish the process once they consider that a good enough overlay has been attained. Hence, skull-face overlay is a very challenging, subjective, error prone, and time consuming part of the whole process. Though the numerical assessment of the method quality has not been achieved yet, computer vision and soft computing arise as powerful tools to automate it, dramatically reducing the time taken by the expert and obtaining an unbiased overlay result. In this manuscript, we justify and analyze the use of these techniques to properly model the skull-face overlay problem. We also present the automatic technical procedure we have developed using these computational methods and show the four overlays obtained in two craniofacial superimposition cases. This automatic procedure can be thus considered as a tool to aid forensic anthropologists to develop the skull-face overlay, automating and avoiding subjectivity of the most tedious task within craniofacial superimposition.

Genetic linkage analysis identifies Pas1 as the common locus modulating lung tumorigenesis and acute inflammatory response in mice.

Selective breeding for the acute inflammatory response (AIR) generated two mouse lines characterized by maximum (AIRmax) and minimum (AIRmin) responses, explained by the additive effect of alleles differentially fixed in quantitative trait loci (QTLs). These mice also differ in their susceptibility to lung tumorigenesis, raising the possibility that the same loci are involved in the control of both phenotypes. To map the QTLs responsible for the different phenotypes, we carried out a genome-wide linkage analysis using single-nucleotide polymorphism arrays in a pedigree consisting of 802 mice, including 693 (AIRmax × AIRmin)F2 intercross mice treated with urethane and phenotyped for AIR and lung tumor multiplicity. We mapped five loci on chromosomes 4, 6, 7, 11 and 13 linked to AIR (logarithm of odds (LOD)=3.56, 3.52, 15.74, 7.74 and 3.34, respectively) and two loci linked to lung tumor multiplicity, on chromosomes 6 and 18 (LOD=12.18 and 4.69, respectively). The known pulmonary adenoma susceptibility 1 (Pas1) locus on chromosome 6 was the only locus linked to both phenotypes, suggesting that alleles of this locus were differentially fixed during breeding and selection of AIR mice. These results represent a step toward understanding the link between inflammation and cancer.

Association study by genetic clustering detects multiple inflammatory response loci in non-inbred mice

We tested the possibility to map loci affecting the acute inflammatory response (AIR) in an (AIRmax AIRmin) F2 intercrossmouse population derived from non-inbred parents, by association analysis in the absence of pedigree information. Using 1064 autosomal single nucleotide polymorphisms (SNPs), we clustered the intercross population into 12 groups of genetically related individuals. Association analysis adjusted for genetic clusters allowed to identify two loci, inflammatory response modulator 1 (Irm1) on chromosome 7 previously detected by genetic linkage analysis in the F2 mice, and a new locus onchromosome 5 (Irm2), linked to the number of infiltrating cells in subcutaneous inflammatory exudates (Irm1: P»6.3 10 7; Irm2: P»8.2 10 5) and interleukin 1 beta (IL-1b) production (Irm1: P»1.9 10 16; Irm2: P»1.1 10 6). Use of a polygenic model based on additive effects of the rare alleles of 15 or 18 SNPs associated at suggestive genome-wide statistical threshold(Po3.4 10 3) with the number of infiltrating cells or IL-1b production, respectively, allowed prediction of the inflammatory response of progenitor AIR mice. Our findings suggest the usefulness of association analysis in combination with genetic clustering to map loci affecting complex phenotypes in non-inbred animal species.

Association study by genetic clustering detects multiple inflammatory response loci in non-inbred mice.

We tested the possibility to map loci affecting the acute inflammatory response (AIR) in an (AIRmax × AIRmin) F2 intercross mouse population derived from non-inbred parents, by association analysis in the absence of pedigree information. Using 1064 autosomal single nucleotide polymorphisms (SNPs), we clustered the intercross population into 12 groups of genetically related individuals. Association analysis adjusted for genetic clusters allowed to identify two loci, inflammatory response modulator 1 (Irm1) on chromosome 7 previously detected by genetic linkage analysis in the F2 mice, and a new locus on chromosome 5 (Irm2), linked to the number of infiltrating cells in subcutaneous inflammatory exudates (Irm1: P=6.3 × 10(-7); Irm2: P=8.2 × 10(-5)) and interleukin 1 beta (IL-1ß) production (Irm1: P=1.9 × 10(-16); Irm2: P=1.1 × 10(-6)). Use of a polygenic model based on additive effects of the rare alleles of 15 or 18 SNPs associated at suggestive genome-wide statistical threshold (P<3.4 × 10(-3)) with the number of infiltrating cells or IL-1ß production, respectively, allowed prediction of the inflammatory response of progenitor AIR mice. Our findings suggest the usefulness of association analysis in combination with genetic clustering to map loci affecting complex phenotypes in non-inbred animal species.

Periodontitis and arthritis interaction in mice involves a shared hyper-inflammatory genotype and functional immunological interferences.

Periodontitis (PD) and rheumatoid arthritis (RA) have been found to be clinically associated and to share the chronic nature of the inflammatory reaction associated with bone resorption activity. However, the mechanisms underlying such association are unknown. Therefore, we examined the basis of Actinobacillus actinomycetemcomitans- and Porphyromonas gingivalis-induced PD and pristane-induced arthritis (PIA) interaction in mice. Higher severity PD in the genetically inflammation prone acute inflammatory reactivity maximum (AIRmax) mice strain was associated with higher levels of TNF-alpha, IL-1beta, IL-17, matrix metalloproteinase (MMP)-13, and RANKL, whereas PD/PIA co-induction resulted in even higher levels of IL-1beta, IFN-gamma, IL-17, RANKL, and MMP-13 levels. Conversely, PD/PIA co-induction in AIRmin strain did not alter the course of both pathologies. PIA/PD co-induction resulted in altered expression of T-cell subsets transcription factors expression, with T-bet and RORgamma levels being upregulated, whereas GATA-3 levels were unaltered. Interestingly, PIA induction resulted in alveolar bone loss, such response being highly dependent on the presence of commensal oral bacteria. No differences were found in PIA severity parameters by PD co-induction. Our results show that the interaction between experimental PD and arthritis in mice involves a shared hyper-inflammatory genotype and functional interferences in innate and adaptive immune responses.

Periodontitis and arthritis interaction in mice involves a shared hyper-inflammatory genotype and functional immunological interferences

Periodontitis (PD) and rheumatoid arthritis (RA) have been found to be clinically associated and to share the chronic nature of the inflammatory reaction associated with bone resorption activity. However, the mechanisms underlying such association areunknown. Therefore, we examined the basis of Actinobacillus actinomycetemcomitans- and Porphyromonas gingivalis-inducedPD and pristane-induced arthritis (PIA) interaction in mice. Higher severity PD in the genetically inflammation prone acute inflammatory reactivity maximum (AIRmax) mice strain was associated with higher levels of TNF-a, IL-1b, IL-17, matrix metalloproteinase (MMP)-13, and RANKL, whereas PD/PIA co-induction resulted in even higher levels of IL-1b, IFN-g, IL-17, RANKL, and MMP-13 levels. Conversely, PD/PIA co-induction in AIRmin strain did not alter the course of both pathologies. PIA/PD co-induction resulted in altered expression of T-cell subsets transcription factors expression, with T-bet and RORg levels being upregulated, whereas GATA-3 levels were unaltered. Interestingly, PIA induction resulted in alveolar bone loss, such response being highly dependent on the presence of commensal oral bacteria. No differences were found in PIA severity parameters by PD co-induction. Our results show that the interaction between experimental PD and arthritis in mice involves a shared hyper-inflammatory genotype and functional interferences in innate and adaptive immune responses.

A new model of outbred genetically selected mice which present a strong acute inflammatory response in the absence of complement component C5.

OBJECTIVE: Mice selected for a strong (AIRmax) or weak (AIRmin) acute inflammatory response present different susceptibilities to bacterial infections, autoimmune diseases and carcinogenesis. Variations in these phenotypes have been also detected in AIRmax and AIRmin mice rendered homozygous for Slc11a1 resistant (R) and susceptible (S) alleles. Our aim was to investigate if the phenotypic differences observed in these mice was related to the complement system. MATERIAL: AIRmax and AIRmin mice and AIRmax and AIRmin groups homozygous for the resistance (R) or susceptibility (S) alleles of the solute carrier family 11a1 member (Slc11a1) gene, formerly designated Nramp-1. METHODS AND RESULTS: While no difference in complement activity was detected in sera from AIRmax and AIRmin strains, all sera from AIRmax Slc11a1 resistant mice (AIRmax(RR)) presented no complement-dependent hemolytic activity. Furthermore, C5 was not found in their sera by immunodiffusion and, polymerase chain reaction and DNA sequencing of its gene demonstrated that AIRmax(RR) mice are homozygous for the C5 deficient (D) mutation previously described in A/J. Therefore, the C5D allele was fixed in homozygosis in AIRmax(RR) line. CONCLUSIONS: The AIRmax(RR) line is a new experimental mouse model in which a strong inflammatory response can be triggered in vivo in the absence of C5.

Slc11a1 (formerly NRAMP1) gene modulates both acute inflammatory reactions and pristane-induced arthritis in mice.

Mice selected for the maximum acute inflammatory reaction (AIRmax) are highly susceptible to pristane-induced arthritis (PIA), whereas mice selected for the minimum response (AIRmin) are resistant. These lines show distinct patterns of leukocyte infiltration and R and S allele frequency disequilibrium of the solute carrier family 11a member 1 (Slc11a1) gene. In order to study the interactions of the Slc11a1 R and S alleles with the inflammation modulating Quantitative Trait Loci (QTL) during PIA development, homozygous AIRmax(RR), AIRmax(SS), AIRmin(RR) and AIRmin(SS) lines were produced by genotype-assisted breedings. These mice received two intraperitoneal injections of 0.5 ml pristane at 60-day intervals, and the subsequent development of arthritis was assessed for 210 days. Cytokine-secreting cell profiles were investigated using enzyme-linked immunospot. Arthritis incidence in AIRmax(RR) mice reached 29%, whereas PIA incidence in AIRmax(SS) mice was 70% by day 180. AIRmin(RR) mice were resistant, whereas 13.3% of AIRmin(SS) mice became arthritic. The presence of the defective S allele also increased arthritis severity, although acute inflammation was higher in mice bearing the R allele. A predominant Th0/Th2-type response in Slc11a1(SS) mice was observed. These results indicate that Slc11a1 is a strong candidate for the QTL modulating acute inflammation and for PIA.

Tumores de la cavidad oral y huesos maxilares en pediatría. Revisión de 122 pacientes / Oral cavity and maxillary bone tumors in pediatrics. Review of 122 patients

Se presenta un estudio retrospectivo de 122 pacientes con tumores que se manifestaron en la cavidad oral, con localización inicial en maxilares o partes blandas (se excluyeron los tumores de la cara sin compromiso bucal). La edad promedio fue 9 años y 6 meses (rango de 1 día a 17 años). El 62 por ciento se presentó en varones. La localización inicial de los tumores fue en hueso en el 53 por ciento de los casos y en partes blandas en el 47 por ciento; 82 pacientes tuvieron lesiones benignas y 40 lesiones malignas. Las manifestaciones al ingreso fueron: tumor palpable o visible (39 por ciento), tumor más dolor (22 por ciento), dolor (19 por ciento) y otros como caída de dientes, parálisis, fiebre o asímetría facial (20 por ciento). La rutina de estudio comprendió radiografía panorámica de maxilar, centellografía ósea (gammacámara con Tecnesio 99), tomografía axial computada (TAC) y resonancia nuclear magnética (RNM). Los pacientes fueron tratados en forma multidisciplinaria siendo la cirugía (punción aspiración con aguja fina, biopsia y/o resección)el procedimiento inicial en la mayoría de ellos. De acuerdo al algoritmo todos los pacientes con lesión ósea fueron estudiados con Rx simple y TAC, 89 por ciento de positividad en ambas, previas a la biopsia por punción. De igual menera en los tumores de partes blandas la TAC mantuvo su utilidad, no así la Rx simple que fue reeplazada por la ecografía cuando se detectó ausencia de compromiso óseo. Las lesiones benignas predominaron (78/122) a nivel de hueso o de partes blandas. la curación en ella fue la regla. En lo que respecta a los tumores, primarios de la región (11/40) correspondieron inicialmente a partes blandas y raramente a hueso, en los que fue frecuente el compromiso metastático o multicéntrico.

Tumores de la cavidad oral y huesos maxilares en pediatría. Revisión de 122 pacientes / Oral cavity and maxillary bone tumors in pediatrics. Review of 122 patients

Se presenta un estudio retrospectivo de 122 pacientes con tumores que se manifestaron en la cavidad oral, con localización inicial en maxilares o partes blandas (se excluyeron los tumores de la cara sin compromiso bucal). La edad promedio fue 9 años y 6 meses (rango de 1 día a 17 años). El 62 por ciento se presentó en varones. La localización inicial de los tumores fue en hueso en el 53 por ciento de los casos y en partes blandas en el 47 por ciento; 82 pacientes tuvieron lesiones benignas y 40 lesiones malignas. Las manifestaciones al ingreso fueron: tumor palpable o visible (39 por ciento), tumor más dolor (22 por ciento), dolor (19 por ciento) y otros como caída de dientes, parálisis, fiebre o asímetría facial (20 por ciento). La rutina de estudio comprendió radiografía panorámica de maxilar, centellografía ósea (gammacámara con Tecnesio 99), tomografía axial computada (TAC) y resonancia nuclear magnética (RNM). Los pacientes fueron tratados en forma multidisciplinaria siendo la cirugía (punción aspiración con aguja fina, biopsia y/o resección)el procedimiento inicial en la mayoría de ellos. De acuerdo al algoritmo todos los pacientes con lesión ósea fueron estudiados con Rx simple y TAC, 89 por ciento de positividad en ambas, previas a la biopsia por punción. De igual menera en los tumores de partes blandas la TAC mantuvo su utilidad, no así la Rx simple que fue reeplazada por la ecografía cuando se detectó ausencia de compromiso óseo. Las lesiones benignas predominaron (78/122) a nivel de hueso o de partes blandas. la curación en ella fue la regla. En lo que respecta a los tumores, primarios de la región (11/40) correspondieron inicialmente a partes blandas y raramente a hueso, en los que fue frecuente el compromiso metastático o multicéntrico.(AU)

The anti-IRBP IgG1 and IgG2a response does not correlate with susceptibility to experimental autoimmune uveitis.

Susceptibility to experimental autoimmune uveitis (EAU) in inbred mice has been associated with a dominant Th1 response. Elevated anti-inter-photoreceptor retinoid-binding protein (anti-IRBP) IgG2a/IgG1 antibody ratios have been implicated as candidate markers to predict disease severity. In the present study, both the anti-IRBP antibody isotype and severity of EAU phenotypes were examined in 4 non-isogenic genetically selected mouse lines to determine if they can be used as general markers of disease. Mice between 8 and 12 weeks old selected for high (H(III)) or low (L(III)) antibody response and for maximum (AIR(MAX)) or minimum (AIR(MIN)) acute inflammatory reaction (AIR) were immunized with IRBP. Each experiment was performed with at least 5 mice per group. EAU was evaluated by histopathology 21 days after immunization and the minimal criterion was inflammatory cell infiltration of the ciliary body, choroid and retina. Serum IgG1- and IgG2a-specific antibodies were determined by ELISA. EAU was graded by histological examination of the enucleated eyes. The incidence of EAU was lower in AIR(MIN) mice whereas in the other strains approximately 40% of the animals developed the disease. Low responder animals did not produce anti-IRBP IgG2a antibodies or interferon-gamma. No correlation was observed between susceptibility to EAU and anti-IRBP isotype profiles. Susceptibility to EAU is related to the intrinsic capacity to mount higher inflammatory reactions and increased production of anti-IRBP IgG2a isotype is not necessarily a marker of this immunologic profile.

The anti-IRBP IgG1 and IgG2a response does not correlate with susceptibility to experimental autoimmune uveitis

Susceptibility to experimental autoimmune uveitis (EAU) in inbred mice has been associated with a dominant Th1 response. Elevated anti-inter-photoreceptor retinoid-binding protein (anti-IRBP) IgG2a/IgG1 antibody ratios have been implicated as candidate markers to predict disease severity. In the present study, both the anti-IRBP antibody isotype and severity of EAU phenotypes were examined in 4 non-isogenic genetically selected mouse lines to determine if they can be used as general markers of disease. Mice between 8 and 12 weeks old selected for high (H III) or low (L III) antibody response and for maximum (AIR MAX) or minimum (AIR MIN) acute inflammatory reaction (AIR) were immunized with IRBP. Each experiment was performed with at least 5 mice per group. EAU was evaluated by histopathology 21 days after immunization and the minimal criterion was inflammatory cell infiltration of the ciliary body, choroid and retina. Serum IgG1- and IgG2a-specific antibodies were determined by ELISA. EAU was graded by histological examination of the enucleated eyes. The incidence of EAU was lower in AIR MIN mice whereas in the other strains approximately 40 percent of the animals developed the disease. Low responder animals did not produce anti-IRBP IgG2a antibodies or interferon-gamma. No correlation was observed between susceptibility to EAU and anti-IRBP isotype profiles. Susceptibility to EAU is related to the intrinsic capacity to mount higher inflammatory reactions and increased production of anti-IRBP IgG2a isotype is not necessarily a marker of this immunologic profile.