Morphological and cytochemical characterization of the peripheral blood cells of farmed streaked prochilod Prochilodus lineatus (Characiformes, Prochilodontidae) / [Caracterização morfológica e citoquímica das células do sangue periférico do curimatã Prochilodus lineatus (Characiformes, Prochilodontidae) em cultivo]

Morphological and cytochemical studies of peripheral blood cells of fish have improved the understanding of their functions and cell types. The present study performed the Morphological and cytochemical analysis of the peripheral blood of Prochilodus lineatus, Characiform native to South America, which has been gaining space in local aquaculture and as a species introduced in Asia. Our analysis provided information on the morphological and cytochemical characteristics of the leukocytes, for the formulation of hypothesis about their role in the immune system of the species. It was found that Prochilodus lineatus has morphological and cytochemical features in common with other fish species, mainly of the Characiformes order. However, we detected the presence of heterophils and PAS positive granulocytes simultaneously with neutrophils. We also found that heterophils and PAS positive granulocytes are very similar, both morphologically and cytochemically.(AU)

O estudo da morfologia e da citoquímica das células do sangue periférico dos peixes tem sido eficaz para o entendimento de suas funções e dos tipos celulares. Este estudo realizou a análise morfológica e citoquímica do sangue periférico de Prochilodus lineatus, caracídeo nativo da América do Sul que vem ganhando espaço na aquicultura local e como espécie introduzida na Ásia. Essa análise forneceu informações sobre a morfologia e as características citoquímicas dos leucócitos, visando a hipóteses sobre suas funções. Verificou-se que estas são semelhantes em vários aspectos a outras espécies, principalmente da ordem Characiformes. No entanto, neste estudo detectou-se a presença dos heterofilos e da célula granulocítica especial, simultaneamente à presença dos neutrófilos. Ainda, foi verificado que os heterofilos e a célula granulocítica especial são muito semelhantes morfológica e citoquimicamente.(AU)

Evidence for X(3872)→J/ψπ

We report the first evidence for X(3872) production in two-photon interactions by tagging either the electron or the positron in the final state, exploring the highly virtual photon region. The search is performed in e^{+}e^{-}→e^{+}e^{-}J/ψπ^{+}π^{-}, using 825 fb^{-1} of data collected by the Belle detector operated at the KEKB e^{+}e^{-} collider. We observe three X(3872) candidates, where the expected background is 0.11±0.10 events, with a significance of 3.2σ. We obtain an estimated value for Γ[over ˜]_{γγ}B(X(3872)→J/ψπ^{+}π^{-}) assuming the Q^{2} dependence predicted by a cc[over ¯] meson model, where -Q^{2} is the invariant mass squared of the virtual photon. No X(3915)→J/ψπ^{+}π^{-} candidates are found.

Use of synthetic oligonucleotides for determination of HTLV-1 proviral load by real-time PCR: a helpful alternative approach in the clinical management.

AIMS: To evaluate the potential use of synthetic oligonucleotides as a standard curve for proviral load (PVL) of human T-cell leukaemia virus type 1 (HTLV-1) quantification in peripheral blood mononuclear cells (PBMC) of HTLV-1-infected individuals by quantitative real-time polymerase chain reaction (qPCR) analysis. METHODS AND RESULTS: Synthetic oligonucleotides based on HTLV-1 genome were customized to use as a standard curve. Twelve anti-HTLV-1-positive samples with known HTLV-1 PVL, previously quantified by qPCR assay using TARL-2 cells as a conventional standard curve, were submitted to the new protocol. The proviral quantification levels had a high concordance with qPCR results using a conventional standard curve. The results demonstrate that the conventional standard curve can be replaced by a synthetic standard curve due to its ability to quantification based on the linearity and qPCR efficiency and similar results with a validated qPCR assay using a conventional standard curve. CONCLUSIONS: Synthetic oligonucleotides standard curves could be a very useful tool on HTLV-1 diagnosis and absolute HTLV-1 PVL quantification. SIGNIFICANCE AND IMPACT OF THE STUDY: HTLV-1 PVL determination using synthetic oligonucleotides standard curve by qPCR could be a helpful alternative for the laboratories that monitor infected patients as an important prognostic factor in HTLV-1-associated diseases progression. Also, it can decrease costs and overcome the biological limitations of the plasmid curve.

Laparoendoscopic Single-site Plus 1-port Donor Nephrectomy: Division of Roles to Shorten Warm Ischemic Time.

BACKGROUND: In this study we present our new surgical procedure, laparoendoscopic single-site surgery plus 1 for donor nephrectomy (LESS+1-DN), which shortens warm ischemic time (WIT) and improves surgical outcomes. METHODS: From January 2013 to February 2017, 15 patients who underwent LESS-DN and 41 patients who underwent LESS+1-DN at our institution were evaluated retrospectively. Patients were divided into 3 groups: group A, 15 cases of LESS-DN; group B, the first 15 patients who underwent LESS+1-DN; and group C, 26 patients who underwent subsequent LESS+1-DN. To reduce WIT, we clearly defined the roles of the surgeon and first assistant in the 26 subsequent LESS+1-DN cases. The surgeon dissected the renal pedicle and harvested the kidney graft using a recovery bag and the first assistant held the recovery bag. RESULTS: The mean operative time in group C (213.7 minutes) was significantly shorter than that in groups A (253.3 minutes) and B (253.8 minutes). The WIT in group C (195.2 seconds) was significantly shorter than that in groups A (389.8 seconds) and B (313.2 seconds). Open conversion was required in 1 case in group A. None of the donors required conversion to open surgery and no perioperative complications occurred in groups B and C. Linear regression analysis of the LESS+1-DN operative times and consecutive case numbers demonstrated a shallow learning curve (R2 = 0.392, P < .05). CONCLUSION: Our new procedure that divides the roles of the operator and the first assistant contributed significantly to a shortening of WIT. Dividing roles can facilitate a safer laparoscopic donor nephrectomy.

Recurrence of Progressive Familial Intrahepatic Cholestasis Type 2 Phenotype After Living-donor Liver Transplantation: A Case Report.

BACKGROUND: Progressive familial intrahepatic cholestasis 2 (PFIC2) is the result of mutations in the ABCB11, which encodes for bile salt export pump (BSEP). An absence of BSEP in the canalicular membrane causes cholestasis and leads to the development of end-stage liver disease in the first decade of life. Liver transplantation (LT) has been considered curative for BSEP disease. However, patients with PFIC2 having undergone LT have recently been reported to develop recurrence of cholestasis together with the clinical and histological features of primary BSEP disease. CASE REPORT: We herein present a rare case of a patient with PFIC2 who developed post-transplantation recurrence of progressive intrahepatic cholestasis due to antibodies against BSEP after living-donor LT, which mimicked primary BSEP disease. The patient had mutations in the ABCB11 gene, resulting in the complete absence of BSEP in the native liver, explaining the lack of tolerance. Immunofluorescence staining of normal human liver sections with the patient's serum and using an anti-human immunoglobulin G antibody to detect serum antibodies showed reactivity to the BSEP epitope in the canalicular membrane. We suggest that the patients having undergone LT had been associated with a risk of autoantibody formation against the BSEP protein. The absence of primary tolerance for the BSEP epitopes may explain the formation of the anti-BSEP antibodies after LDLT.

Radiation track, DNA damage and response-a review.

The purpose of this paper has been to review the current status and progress of the field of radiation biophysics, and draw attention to the fact that physics, in general, and radiation physics in particular, with the aid of mathematical modeling, can help elucidate biological mechanisms and cancer therapies. We hypothesize that concepts of condensed-matter physics along with the new genomic knowledge and technologies and mechanistic mathematical modeling in conjunction with advances in experimental DNA (Deoxyrinonucleic acid molecule) repair and cell signaling have now provided us with unprecedented opportunities in radiation biophysics to address problems in targeted cancer therapy, and genetic risk estimation in humans. Obviously, one is not dealing with 'low-hanging fruit', but it will be a major scientific achievement if it becomes possible to state, in another decade or so, that we can link mechanistically the stages between the initial radiation-induced DNA damage; in particular, at doses of radiation less than 2 Gy and with structural changes in genomic DNA as a precursor to cell inactivation and/or mutations leading to genetic diseases. The paper presents recent development in the physics of radiation track structure contained in the computer code system KURBUC, in particular for low-energy electrons in the condensed phase of water for which we provide a comprehensive discussion of the dielectric response function approach. The state-of-the-art in the simulation of proton and carbon ion tracks in the Bragg peak region is also presented. The paper presents a critical discussion of the models used for elastic scattering, and the validity of the trajectory approach in low-electron transport. Brief discussions of mechanistic and quantitative aspects of microdosimetry, DNA damage and DNA repair are also included as developed by the authors' work.

Impact of Donor-Specific Antibodies on Graft Fibrosis After Pediatric Living Donor Liver Transplantation for Biliary Atresia.

BACKGROUND: Pediatric living donor liver transplant (LDLT) patients sometimes develop graft fibrosis after non-recurrent diseases such as biliary atresia (BA). Donor-specific antibodies (DSA) have recently been shown to play a possible role in graft damage after liver transplantation. We report the impact of DSA on pediatric LDLT for BA patients. METHODS: Patients under age 18 years who received LDLT for BA at our institution and who had at least 5 years' follow-up were identified, and 23 were eventually enrolled in this study. Pathological findings were assessed with the use of the last available biopsy. Patients were divided into 2 groups, DSA-positive and DSA-negative. Graft fibrosis after LDLT was assessed according to DSA groups. RESULTS: The mean patient age at transplant was 2.6 years. The mean time to the last available biopsy after LDLT was 8.2 years (4.8-15.6 years); 6 patients (26%) showed no fibrosis, whereas fibrosis was graded as F1, F2, or F3 in 8 patients (35%), 8 patients (35%), and 1 patient, respectively. DSA were observed in 12 patients (52%). Moderate graft fibrosis (F2 and F3) was found in 7 (58%) of the DSA-positive group, but only 2 (18%) of the DSA-negative group, showing a statistically significant difference (P < .05). Pre-transplant cross-matching was performed in 17 patients. The 2 patients with a positive cross-match were DSA-positive. Six cross-match-negative patients developed de novo DSA after LDLT. CONCLUSIONS: Graft fibrosis was observed after LDLT for BA during long-term follow-up, more commonly in DSA-positive patients. DSA may play a role in fibrosis formation.

Disseminated Metastatic Tissue Calcification After Orthotopic Liver Transplantation: A Case Report.

BACKGROUND: Hypercalcemia has been observed in patients after liver transplantation. However, it is rare that the hypercalcemia induced disseminated tissue calcification and heart failure. CASE REPORT: We report a rare case of heart failure caused by disseminated metastatic tissue calcification that involved extensive progressive myocardial calcification after liver transplantation. A 20-year-old man with end-stage liver disease due to biliary atresia underwent ABO-incompatible living donor liver transplantation. After successful transplantation, he suffered from antibody-mediated rejection. Subsequently, ABO-matched cadaveric liver retransplantation was successfully performed. Hypercalcemia developed gradually following the second transplantation. His serum calcium level increased to 18.3 mg/dL with sudden onset of ventricular tachycardia. Although he was resuscitated with a cardiopulmonary support device, he died of heart and liver failure. Histopathologic examination revealed systemic disseminated metastatic tissue calcification, including massive myocardial calcification. CONCLUSION: Progressive worsening of hypercalcemia resulted in disseminated metastatic tissue calcification and massive metastatic myocardial calcification, which led to heart failure after liver transplantation. Because hypercalcemia after liver transplantation can cause fatal tissue calcification, early intervention for hypercalcemia should be considered.

Estimation of the width of free margin with a significant impact on local recurrence in surgical resection of oral squamous cell carcinoma.

The purpose of this study was to estimate the width of free margin with a significant impact on local recurrence in surgical resection of oral squamous cell carcinoma (OSCC). Clinical and pathological data of 127 consecutive patients who underwent radical resection of OSCC were analyzed retrospectively. The local control rate was compared between patients with clear, close, and involved surgical margins, changing the required width of free margin for the definition of 'close surgical margin' (from 1 to 5mm). If a free margin of within 1, 2, or 4mm was judged a close margin, the risk of local recurrence was significantly different among the patients with clear, close, and involved surgical margins. If the definition of close margin was within 5mm of the resection margin, the difference between clear and close margin did not reach statistical significance. The results of this study suggest that 5mm of clearance at the surgical resection margin should be the index of oncological surgery. More than 5mm of histological free margin around OSCC is not justified in terms of the risk management of local recurrence and the resultant morbidity.

Microdosimetry of the full slowing down of protons using Monte Carlo track structure simulations.

The article investigates two approaches in microdosimetric calculations based on Monte Carlo track structure (MCTS) simulations of a 160-MeV proton beam. In the first approach, microdosimetric parameters of the proton beam were obtained using the weighted sum of proton energy distributions and microdosimetric parameters of proton track segments (TSMs). In the second approach, phase spaces of energy depositions obtained using MCTS simulations in the full slowing down (FSD) mode were used for the microdosimetric calculations. Targets of interest were water cylinders of 2.3-100 nm in diameters and heights. Frequency-averaged lineal energies ([Formula: see text]) obtained using both approaches agreed within the statistical uncertainties. Discrepancies beyond this level were observed for dose-averaged lineal energies ([Formula: see text]) towards the Bragg peak region due to the small number of proton energies used in the TSM approach and different energy deposition patterns in the TSM and FSD of protons.

First molar cross-bite is more closely associated with a reverse chewing cycle than anterior or pre-molar cross-bite during mastication.

A posterior cross-bite is defined as an abnormal bucco-lingual relationship between opposing molars, pre-molars or both in centric occlusion. Although it has been reported that patients with unilateral posterior cross-bite often show unique chewing patterns, the relationship between the form of cross-bite and masticatory jaw movement remains unclear in adult patients. The objective of this study was to investigate masticatory jaw movement among different forms of cross-bite. One hundred and one adults were recruited in this study: 27 had unilateral first molar cross-bite (MC group); 28, unilateral pre-molar cross-bite (PC group); 23, anterior cross-bite (AC group); and 23, normal occlusion (control group). Masticatory jaw movement of the lower incisor point was recorded with six degrees of freedom jaw-tracking system during unilateral mastication. Our results showed that the reverse chewing ratio during deliberate unilateral mastication was significantly larger in the MC group than in the PA (P < 0.001), AC (P < 0.001) and control (P < 0.001) groups. These findings suggest that compared to the anterior or pre-molar cross-bite, the first molar cross-bite is more closely associated with a higher prevalence of a reverse chewing cycle.

Posterior scissors-bite: masticatory jaw movement and muscle activity.

Scissors-bite is a malocclusion characterised by buccal inclination or buccoversion of the maxillary posterior tooth and/or linguoclination or linguoversion of the mandibular posterior tooth. This type of malocclusion causes reduced contact of the occlusal surfaces and can cause excessive vertical overlapping of the posterior teeth. This case-control study is the first to evaluate both masticatory jaw movement and masseter and temporalis muscle activity in patients with unilateral posterior scissors-bite. Jaw movement variables and surface electromyography data were recorded in 30 adult patients with unilateral posterior scissors-bite malocclusion and 18 subjects with normal occlusion in a case-control study. The chewing pattern on the scissors-bite side significantly differed from that of the non-scissors-bite side in the patients and of the right side in the normal subjects. These differences included a narrower chewing pattern (closing angle, P < 0.01; cycle width, P < 0.01), a longer closing duration (P < 0.05), a slower closing velocity (P < 0.01) and lower activities of both the temporalis (P < 0.05) and the masseter (P < 0.05) muscles on the working side. In 96% of the patients with unilateral posterior scissors-bite, the preferred chewing side was the non-scissors-bite side (P = 0.005). These findings suggest that scissors-bite malocclusion is associated with the masticatory chewing pattern and muscle activity, involving the choice of the preferred chewing side in patients with unilateral posterior scissors-bite.

Influence of maximum bite force on jaw movement during gummy jelly mastication.

It is known that maximum bite force has various influences on chewing function; however, there have not been studies in which the relationships between maximum bite force and masticatory jaw movement have been clarified. The aim of this study was to investigate the effect of maximum bite force on masticatory jaw movement in subjects with normal occlusion. Thirty young adults (22 men and 8 women; mean age, 22.6 years) with good occlusion were divided into two groups based on whether they had a relatively high or low maximum bite force according to the median. The maximum bite force was determined according to the Dental Prescale System using pressure-sensitive sheets. Jaw movement during mastication of hard gummy jelly (each 5.5 g) on the preferred chewing side was recorded using a six degrees of freedom jaw movement recording system. The motion of the lower incisal point of the mandible was computed, and the mean values of 10 cycles (cycles 2-11) were calculated. A masticatory performance test was conducted using gummy jelly. Subjects with a lower maximum bite force showed increased maximum lateral amplitude, closing distance, width and closing angle; wider masticatory jaw movement; and significantly lower masticatory performance. However, no differences in the maximum vertical or maximum anteroposterior amplitudes were observed between the groups. Although other factors, such as individual morphology, may influence masticatory jaw movement, our results suggest that subjects with a lower maximum bite force show increased lateral jaw motion during mastication.

Conversion to prolonged-release tacrolimus for pediatric living related donor liver transplant recipients.

Prolonged-release tacrolimus allows for once-daily dosing. Although many adult recipients have been switched from standard tacrolimus, prolonged-release tacrolimus has not been popular for pediatric patients despite the potential benefits for medication compliance. We report on prolonged-release tacrolimus for 11 pediatric living related donor liver transplant (LRDLT) recipients. Patients under 18 years of age who were receiving standard tacrolimus-based immunosuppression and steroid taper underwent conversion from standard to prolonged-release tacrolimus. We monitored tacrolimus trough levels and liver function tests (LFTs). We also assessed adverse effects and satisfaction levels for prolonged-release tacrolimus. Mean age at transplantation was 4.3 years. The mean duration of follow-up was 12 months. The ratios of trough levels with prolonged-release vs standard tacrolimus were 0.97, 0.95, and 0.92 at 1, 2, and 4 weeks post conversion, respectively. Two patients discontinued prolonged-release tacrolimus owing to abnormal LFTs and neurological abnormalities, respectively; but symptoms resolved after reconversion. One patient returned to standard tacrolimus and the other was converted to cyclosporine. Once-daily administration satisfied 89% of patients. In the overall assessment, conversion to prolonged-release tacrolimus satisfied all patients. Prolonged-release tacrolimus was useful for pediatric patients after LRDLT. Trough levels after conversion were compatible with those before conversion. Most patients were satisfied with prolonged-release tacrolimus. However, some patients failed conversion because of unexpected responses. Close observation after conversion is required even if patients have previously had an uneventful course on standard tacrolimus.

Association of uricemia with biochemical and dietary factors in human adults with metabolic syndrome genotyped to C677T polymorphism in the methylenetetrahydrofolate reductase gene.

It is suggested that hyperuricemia is a marker of cardiovascular risk in human adults with metabolic syndrome (MS). The C677T polymorphism in the gene encoding the enzyme methylenetetrahydrofolate reductase (MTHFR) is associated with hyperuricemia. Data on factors associated with uricemia in human adults with MS genotyped for this polymorphism are lacking. We aimed to investigate the factors associated with uricemia in human adults with MS genotyped for the C677T polymorphism in the MTHFR gene. Cross-sectional study was conducted with 63 human adults (24 men and 39 women) with MS. Body weight, body mass index, waist circumference, body fat, glycemia, lipid profile, uricemia, insulinemia, homocysteinemia, plasma folate, erythrocyte folate, blood pressure, smoking, diuretics use, usual dietary alcohol and protein intakes, MTHFR and the presence of the C677T polymorphism in the gene were assessed. Hyperuricemia was observed in 16 (25.4%) human adults (10 men and 6 women). In the group, 33% (n = 21) showed the C677T polymorphism, being 19 heterozygous and 2 mutant homozygous. A significant association between hyperuricemia and C677T polymorphism was not verified. Uricemia was positively associated with homocys-teinemia (r = 0.43, p < 0.05), triglyceridemia (r = 0.41, p<0.05), serum concentrations of very-low-density lipoprotein (r = 0.27, p< 0.05) and the habitual alcohol intake (r = 0.37, p < 0.05). However, only homocysteinemia, triglyc-eridemia, and habitual alcohol intake remained in the final model of linear regression. In human adults with MS geno-typed for the C677T polymorphism in the MTHFR gene, uricemia was positively associated with homocysteinemia, triglyceridemia and the habitual alcohol intake.

A database of frequency distributions of energy depositions in small-size targets by electrons and ions.

Linear energy transfer (LET) is an average quantity, which cannot display the stochastics of the interactions of radiation tracks in the target volume. For this reason, microdosimetry distributions have been defined to overcome the LET shortcomings. In this paper, model calculations of frequency distributions for energy depositions in nanometre size targets, diameters 1-100 nm, and for a 1 µm diameter wall-less TEPC, for electrons, protons, alpha particles and carbon ions are reported. Frequency distributions for energy depositions in small-size targets with dimensions similar to those of biological molecules are useful for modelling and calculations of DNA damage. Monte Carlo track structure codes KURBUC and PITS99 were used to generate tracks of primary electrons 10 eV to 1 MeV, and ions 1 keV µm(-1) to 300 MeV µm(-1) energies. Distribution of absolute frequencies of energy depositions in volumes with diameters of 1-100 nm randomly positioned in unit density water irradiated with 1 Gy of the given radiation was obtained. Data are presented for frequency of energy depositions and microdosimetry quantities including mean lineal energy, dose mean lineal energy, frequency mean specific energy and dose mean specific energy. The modelling and calculations presented in this work are useful for characterisation of the quality of radiation beam in biophysical studies and in radiation therapy.

[Influence of fat diet in glicidic metabolism in obese women with Pro12Pro genotype in PPARgamma2 gene]. / Influencia de la grasa de la dieta en el metabolismo glucídico de mujeres obesas con el genotipo Pro12Pro en el gen PPARgama2.

INTRODUCTION: The peroxisome proliferator-activated receptor gamma 2 (PPARgamma2) is an adipogenic transcription factor that influences in insulin resistance (IR) in the presence of agonists such as polyunsaturated fatty acids (PUFA). OBJECTIVE: Evaluate the influence of dietary fat in glicidic metabolism in morbidly obese women with Pro12Pro genotype in the gene PPARgamma2. METHODS: Were selected 25 women with genotype Pro12Pro. The fat intake was estimated by food records, being used for the division of groups, GA (until 30% of the total energy expenditure (TEE)) and GB (greater than 30% of the TEE). Biochemical and anthropometric evaluations were conducted in fasting, following the test meal high in n-6 PUFA and postprandial biochemical evaluations. IR and insulin sensitivity (IS) were assessed by HOMA-IR (Homeostasis Model Assessment) and QUICKI (Quantitative Insulin Sensitivity Check Index), respectively. RESULTS AND DISCUSSION: GA presented normal HOMAIR and QUICKI. GB presented higher body mass index (BMI), HOMA-IR, saturated fatty acids (SFA) and monounsaturated (MUFA) intake higher, compared with GA (p < 0.05). In GA, the MUFA intake was negatively correlated with HOMA-IR, fasting glucose and insulin, and positively with QUICKI. The fat and SFA intake contributed to the increase in body mass and IR. However, MUFA intake may have reduced the impact of high fat diet in glicidic metabolism. It is suggested that obese women with Pro12Pro genotype in the PPARgamma2 gene avoid high fat and SFA diets, prioritizing MUFA for controlling obesity and improving the IS.

Observation of a charmoniumlike enhancement in the gammagamma-->omega(J)/psi process.

We report the results of a search for a charmoniumlike state produced in the process gammagamma-->omegaJ/psi in the 3.9-4.2 GeV/c{2} mass region. We observe a significant enhancement, which is well described by a resonant shape with mass M=(3915+/-3+/-2) MeV/c{2} and total width Gamma=(17+/-10+/-3) MeV. This enhancement may be related to one or more of the three charmoniumlike states so far reported in the 3.90-3.95 GeV/c{2} mass region.

The consideration of biological effectiveness of low energy protons using biophysical modeling of the effects induced by exposure of V79 cells.

We have applied Monte Carlo track structure simulations to estimate relative biological effectiveness (RBE) of low-energy protons using biophysical modelling of radiation effects induced by exposure of V79 cells growing in mono-layer. The microscopic energy deposition in cell nucleus and sub-nucleus volumes was investigated in order to understand the reasons of enhanced biological effectiveness near Bragg peak. Theoretical estimations of RBE based on frequency/dose average lineal energy and calculated yields of initial DNA breaks were collated with experimental RBE(M) data. It was found: 1) dose average lineal energy for whole cell nucleus as a function of proton energy shows a distinct peak at 550 keV; 2) the peak values for sub-nucleus volumes are large compared with the whole cell nucleus; 3) the yield of complex DNA breaks correlates with experimental RBE(M) data.

Complement independent antibody-mediated endarteritis and transplant arteriopathy in mice.

Complement fixation, as evidenced by C4d in the microvasculature, is a widely accepted criterion of antibody-mediated rejection. Complement fixation has been shown to be essential in acute antibody-mediated rejection, but its role in chronic rejection has not been addressed. Previous studies showed that passive transfer of complement fixing monoclonal IgG2a anti-H-2Kk into B6.RAG1-/- KO recipients of B10.BR hearts led to progressive chronic transplant arteriopathy (CTA) over 14-28 days, accompanied by C4d deposition. The present studies were designed to test whether complement was required for these lesions. We report that a noncomplement fixing donor-specific alloantibody (DSA, monoclonal IgG1 anti-H-2Kk) injected into B6.RAG1-/- KO recipients of B10.BR hearts also promotes CTA, without C4d deposition. Furthermore, a passive transfer of DSA (monoclonal IgG2a anti-H-2Kk) initiated endarteritis followed by CTA in B6.RAG1-/- mice genetically deficient in the third component of complement (RAG1-/-C3-/-). These studies indicate that antibody to class I MHC antigens can trigger chronic arterial lesions in vivo without complement participation, in contrast to acute antibody-mediated rejection. This pathway may be relevant to C4d-negative chronic rejection sometimes observed in patients with DSA, and argues that lack of C4d deposition does not exclude antibody-mediated chronic rejection.