Systematic and Comparative Analysis of the Burden of Alzheimer´s Disease and Other Dementias in Mexico. Results at the National and Subnational Levels, 1990-2019.

INTRODUCTION: Dementias, including Alzheimer´s disease (AD), are one of the leading causes of disability and mortality in older people. It is a growing health problem in low- and middle-income countries, where epidemiological information is scarce and deficient. The aim of this study was to analyze the burden of AD and other dementias in Mexico from 1990 to 2019 by sex, subnational level, and age groups. METHODS: A secondary analysis was conducted using data from the 2019 Global Burden of Disease, Injury, and Risk Factors Study (GBD). Data on prevalence, incidence, mortality, years of life lost (YLL), years lived with disability (YLD), and disability-adjusted life years (DALY) due to AD and other dementias were obtained. A joinpoint regression analysis was performed to describe the changes in the trend of age-standardized DALY rates by AD and other dementias during the analysis period. RESULTS: AD and other dementias ranked second among neurological disorders producing the most DALY in Mexico. Between 1990 and 2019, prevalence and incidence increased by almost 203%. In 2019, the age-standardized rate per 100,000 inhabitants was: 512 for prevalence, 79.3 for incidence, 73.3 for YLD, 256.9 for YLL and 272.2 for DALY. Likewise, five states concentrated 39% of AD and other dementias cases: Ciudad de México, Estado de México, Veracruz, Jalisco and Puebla. Differences were also observed by sex and age groups. DISCUSSION: Given that the number of older adults in Mexico will significantly rise over the next few decades, AD and other dementias represent one of the most important health challenges. The fact that epidemiological and demographic transformations take place in Mexico in a very diverse way makes it difficult for the country to adequately plan for the growing demands of both people with AD and other dementias and their families.

Impact of a Maternal Motivational Interviewing on Oral Health in the Mother-Child Dyad.

Motivational Interviewing (MI) has been included in dentistry programs. There exists a need for interventions in the mother-child dyad. The aim of this paper was to compare the effects of a MI-based educational program on oral care knowledge, attitudes, and practices (KAP) in the mother-child dyad to a Traditional Education-based program (TE). A community intervention trial was carried out. The experimental and control groups were made up of women between 18 and 45 years of age in the sixth month of gestation. Both groups were provided with TE. The experimental group additionally received a session based on the principles of the MI. Socio-demographic data, children's oral health KAP (COHKAP), that of the mother (MOHKAP), and maternal self-efficacy (MSE) in relation to children's oral health (COHMSE) were recorded. A baseline measurement was made, as well as a six-month follow up. The participants included 135 women with an average age of 24.88 ± 6.00. After intervention, the experimental group's COHKAP, MOHKAP, and COHMSE all increased (p < 0.001). When MI-based interventions are combined with TE, MSE and dental care KAP for the mother-child dyad in pregnant women can be improved.

Effect of soccer footwear on landing mechanics.

Lower-extremity injury is common in soccer. A number of studies have begun to assess why specific lower-extremity injuries occur. However, currently few studies have examined how footwear affects lower-extremity mechanics. In order to address this question, 14 male (age: 22.1 ± 3.9 years, height: 1.77 ± 0.06 m, and mass: 73.3 ± 11.5 kg) and 14 female (age: 22.8 ± 3.1 years, height: 1.68 ± 0.07 m and mass: 64.4 ± 9.2 kg) competitive soccer players underwent a motion analysis assessment while performing a jump heading task. Each subject performed the task in three different footwear conditions (running shoe, bladed cleat, and turf shoe). Two-way analyses of variance were used to examine statistical differences in landing mechanics between the footwear conditions while controlling for gender differences. These comparisons were made during two different parts (prior to and following) of a soccer-specific jump heading task. A statistically significant interaction for the peak dorsiflexion angle (P = 0.02) and peak knee flexion angle (P = 0.05) was observed. Male soccer players exhibited a degree increase in dorsiflexion in the bladed cleat while female soccer players exhibited a three-degree reduction in peak knee flexion in the bladed cleat condition. Other main effects for gender and footwear were also observed. The results suggest that landing mechanics differ based upon gender, footwear, and the type of landing. Therefore, training interventions aimed at reducing lower-extremity injury should consider utilizing sport-specific footwear when assessing movement patterns.

Bronchoscopic lung-volume reduction with Exhale airway stents for emphysema (EASE trial): randomised, sham-controlled, multicentre trial.

BACKGROUND: Airway bypass is a bronchoscopic lung-volume reduction procedure for emphysema whereby transbronchial passages into the lung are created to release trapped air, supported with paclitaxel-coated stents to ease the mechanics of breathing. The aim of the EASE (Exhale airway stents for emphysema) trial was to evaluate safety and efficacy of airway bypass in people with severe homogeneous emphysema. METHODS: We undertook a randomised, double-blind, sham-controlled study in 38 specialist respiratory centres worldwide. We recruited 315 patients who had severe hyperinflation (ratio of residual volume [RV] to total lung capacity of ≥0·65). By computer using a random number generator, we randomly allocated participants (in a 2:1 ratio) to either airway bypass (n=208) or sham control (107). We divided investigators into team A (masked), who completed pre-procedure and post-procedure assessments, and team B (unmasked), who only did bronchoscopies without further interaction with patients. Participants were followed up for 12 months. The 6-month co-primary efficacy endpoint required 12% or greater improvement in forced vital capacity (FVC) and 1 point or greater decrease in the modified Medical Research Council dyspnoea score from baseline. The composite primary safety endpoint incorporated five severe adverse events. We did Bayesian analysis to show the posterior probability that airway bypass was superior to sham control (success threshold, 0·965). Analysis was by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00391612. FINDINGS: All recruited patients were included in the analysis. At 6 months, no difference between treatment arms was noted with respect to the co-primary efficacy endpoint (30 of 208 for airway bypass vs 12 of 107 for sham control; posterior probability 0·749, below the Bayesian success threshold of 0·965). The 6-month composite primary safety endpoint was 14·4% (30 of 208) for airway bypass versus 11·2% (12 of 107) for sham control (judged non-inferior, with a posterior probability of 1·00 [Bayesian success threshold >0·95]). INTERPRETATION: Although our findings showed safety and transient improvements, no sustainable benefit was recorded with airway bypass in patients with severe homogeneous emphysema. FUNDING: Broncus Technologies.

CD28-B7-mediated T cell costimulation in chronic cardiac allograft rejection: differential role of B7-1 in initiation versus progression of graft arteriosclerosis.

Provision of adequate T cell costimulation is critical for the development of acute and chronic allograft rejection. We have previously reported that early blockade of CD28-B7 T cell costimulation prevents the development of graft arteriosclerosis, in the LEW into F344 rat cardiac transplant model. In this study, we used the same model to examine the requirement for CD28-B7-mediated T cell costimulation in the progression of established chronic rejection and examined the individual roles of B7-1 (CD80) and B7-2 (CD86) costimulatory molecules. Late blockade of CD28-B7 T cell costimulation by the fusion protein CTLA4Ig, which binds both CD80 and CD86, attenuated the development of transplant arteriosclerosis, mononuclear cell infiltration, and parenchymal fibrosis in this model. Selective blockade of CD80 using the mutant fusion protein Y100F was as effective as CTLA4Ig in this regard. In contrast to CTLA4Ig, blockade of CD80 alone by Y100F was ineffective at preventing early graft loss and prolonging graft survival when given early after transplantation. This study is the first to demonstrate that late blockade of CD28-B7 T cell costimulation interrupts chronic cardiac allograft rejection, and it indicates the importance of continued T cell activation in this process. This study further defines functional differences between CD80 and CD86 costimulatory molecules in vivo.

1-Aminomethylisoquinoline-4-carboxylates as novel dipeptidylpeptidase IV inhibitors.

Structure-activity relationship within a series of 1-aminoalkylisoquinoline-4-carboxylates as inhibitors of DPP-IV is described. A primary aminomethyl group is required to maintain biological activity. Substitution of the isoquinoline at the 6- and 8-positions with methoxy groups increases potency to 53 times that of the lead compound SDZ 029-576.

Attenuated cardiac allograft vasculopathy in mice with targeted deletion of the transcription factor STAT4.

BACKGROUND: To study transcription factor signaling pathways that mediate cardiac allograft vasculopathy, we used mice with targeted gene deletion of signal transducer and activator of transcription (STAT)4 and STAT6 as recipients in our mouse cardiac transplant model of chronic rejection. METHODS AND RESULTS: At day 55 after transplantation, cardiac grafts placed into STAT4 -/- (n=10) had reduced frequency (24+/-2%) and severity (9+/-4%) of vascular occlusion compared with wild-type controls (n=7, frequency 70+/-12% [P<0.001], severity 25+/-6% [P<0.05]). This decrease was associated with reduced intragraft expression ((32)P RT-PCR and immunohistochemistry) of the Th1 signature cytokines interferon-gamma (P<0.001) and interleukin (IL)-2 (P<0.001). Furthermore, cardiac grafts in STAT4 -/- had fewer infiltrating CD45(+) mononuclear cells (99+/-27 cells/mm(3) compared with 551+/-168 cells/mm(3) in wild-type controls [P<0.05]) and reduced expression of P-selectin (P<0.001) and E-selectin (P<0.01) ligand, recently shown to regulate Th1 cell recruitment. In contrast, in grafts placed into STAT6 -/- (n=11), the development of cardiac allograft vasculopathy (frequency 62+/-8%, severity 28+/-6%) and Th2 cytokine profiles (IL-4, IL-10) were comparable to those in wild-type controls. CONCLUSIONS: Hence, we show that immune responses mediated by STAT4, but not STAT6, contribute to the development of cardiac allograft vasculopathy. We speculate that when present, STAT4-mediated signaling pathways may promote cardiac allograft vasculopathy by directing Th1-specific lymphocyte recruitment, activation, and effector functions.

Cardiac allograft vasculopathy--a changing perspective.

Together, recent clinical and basic studies show that Cardiac allograft vasculopathy (CAV) is multifactorial and regulated by interacting forces at different stages. Data show that at least three forms of vasculopathy (fibrosis, arteritis, atheroma) contribute to the thickening of graft vascular disease. Current therapy to prevent or treat CAV should focus on control of both alloantigen dependent and independent forces. In the future, therapies will be targeted at manipulation of the donor (to reduce antigenicity and ischemic injury) as well as manipulation of the recipient to attenuate or permanently interrupt the immune responses in a donor specific manner.

Donor and recipient contributions of ICAM-1 and P-selectin in parenchymal rejection and graft arteriosclerosis: insights from double knockout mice.

BACKGROUND: Mice with target gene deletions were used in an immunosuppressed, heterotopic mouse cardiac transplant model to investigate the effects of simultaneous deficiencies of ICAM-1 and P-selectin on late cardiac rejection. METHODS: To determine the contribution of donor sources of ICAM-1 and P-selectin, ICAM-1/P-selectin gene deficient (I/P -/-) (n = 7) or wild type (n = 6) donor hearts were placed into CBA recipients. To study recipient sources of ICAM-1 and P-selectin, wild type donor hearts were placed into I/P -/- (n = 7) or wild type (n = 13) recipients. Recipients received a 30-day course of anti-CD4/8 mAb. RESULTS: I/P -/- donor allografts had prolonged survival (52-57 days) compared with wild type allografts (49-51 days). I/P -/- donor allografts underwent parenchymal rejection with mononuclear cell infiltration and developed alpha-smooth muscle actin positive vascular thickening (30 +/- 7% luminal occlusion, n = 78 vessels). Wild type allografts had parenchymal rejection with vascular medial necrosis and an absence of arteriosclerotic thickening (10 +/- 8%, n = 75, p = 0.008). Using the reverse combination, allografts from I/P -/- or wild type recipients had similar graft survival (50-57 days), comparable but variable degrees of parenchymal rejection, and comparable vascular occlusion (22 +/- 15% vs 28 +/- 19%, p = 0.442). CONCLUSION: We have shown that donor and recipient sources of ICAM-1 and P-selectin may have independent roles in leukocyte trafficking to the graft. Simultaneous interruption of donor ICAM-1 and P-selectin delays onset of parenchymal rejection. However, donor I/P deficiency permits arteriosclerotic development, perhaps by attenuating the alloimmune injury. In contrast, recipient deficiency alone does not altergraft outcomes suggesting that the donor is the critical site of ICAM-1 and P-selectin.

NVP-DPP728 (1-[[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(S)- pyrrolidine), a slow-binding inhibitor of dipeptidyl peptidase IV.

Inhibition of dipeptidyl peptidase IV (DPP-IV) has been proposed recently as a therapeutic approach to the treatment of type 2 diabetes. N-Substituted-glycyl-2-cyanopyrrolidide compounds, typified by NVP-DPP728 (1-[[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(S )-p yrrolidine), inhibit degradation of glucagon-like peptide-1 (GLP-1) and thereby potentiate insulin release in response to glucose-containing meals. In the present study NVP-DPP728 was found to inhibit human DPP-IV amidolytic activity with a K(i) of 11 nM, a k(on) value of 1.3 x 10(5) M(-)(1) s(-)(1), and a k(off) of 1.3 x 10(-)(3) s(-)(1). Purified bovine kidney DPP-IV bound 1 mol/mol [(14)C]-NVP-DPP728 with high affinity (12 nM K(d)). The dissociation constant, k(off), was 1.0 x 10(-)(3) and 1.6 x 10(-)(3) s(-)(1) in the presence of 0 and 200 microM H-Gly-Pro-AMC, respectively (dissociation t(1/2) approximately 10 min). Through kinetic evaluation of DPP-IV inhibition by the D-antipode, des-cyano, and amide analogues of NVP-DPP728, it was determined that the nitrile functionality at the 2-pyrrolidine position is required, in the L-configuration, for maximal activity (K(i) of 11 nM vs K(i) values of 5.6 to >300 microM for the other analogues tested). Surprisingly, it was found that the D-antipode, despite being approximately 500-fold less potent than NVP-DPP728, displayed identical dissociation kinetics (k(off) of 1.5 x 10(-)(3) s(-)(1)). NVP-DPP728 inhibited DPP-IV in a manner consistent with a two-step inhibition mechanism. Taken together, these data suggest that NVP-DPP728 inhibits DPP-IV through formation of a novel, reversible, nitrile-dependent complex with transition state characteristics.

Clinical rejection is distinguished from subclinical rejection by increased infiltration by a population of activated macrophages.

It has been reported previously that one-third of protocol renal biopsies in asymptomatic, biochemically stable renal transplant recipients in the first 6 mo show unsuspected subclinical graft rejection (both infiltrate and tubulitis) and that subclinical rejection is a risk factor for chronic renal dysfunction. This study was performed to determine whether differences in phenotype or activation status of graft-infiltrating cells underlie these different manifestations of acute rejection. Biopsies with normal histology (n = 10), subclinical rejection (n = 13), and clinical rejection (n = 9) were studied using immunohistochemistry and computerized image analysis. Subclinical and clinical rejections had similar histologic Banff scores. Univariate analysis showed a trend for a higher infiltration with CD8+ (P = 0.053) and CD68+(P = 0.06) cells in clinical rejection. Of the activation markers studied (CD25, perforin, tumor necrosis factor-alpha), only allograft inflammatory factor-1+-activated macrophages were significantly (P = 0.014) increased in the infiltrate of clinical rejection biopsies. These data suggest that activated macrophages or their products are responsible for acute renal dysfunction associated with clinical rejection episodes.

Alloimmune-mediated apoptosis: comparison in mouse models of acute and chronic cardiac rejection.

BACKGROUND: The purpose of the present study was (1) to compare apoptotic activity in models of acute and chronic rejection and (2) to study the cellular distribution of parenchymal versus inflammatory cell apoptosis. METHODS: Heterotopic cardiac mouse transplantation (CBA into C57BL/6) was used to produce allografts undergoing acute (day 7, untreated recipients, n=6) or chronic (day 55, anti-CD4/8 for 28 days, n=6) rejection. As references, we used 55-day isograft controls (n=5) and native hearts (n=6). To assess apoptotic activity, we quantified DNA laddering (32P incorporation), DNA fragmentation (antinucleosome ELISA), and caspase-1 transcript levels (32P-reverse transcriptase-polymerase chain reaction). To localize apoptosis, we performed terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling. RESULTS: DNA laddering and nucleosome levels were increased in allografts undergoing acute or chronic rejection when compared with both controls. Both parameters were twofold higher in acutely compared with chronically rejecting hearts. Caspase-1 transcript levels were increased in acutely (P<0.0001) and chronically rejecting hearts (P=0.004). Acutely rejecting grafts had more terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive nuclei (53+/-3 nuclei/high-powered field) than chronically rejecting grafts (9+/-1 nuclei/high-powered field, P<0.0001), but the distribution between graft-infiltrating inflammatory cells and myocytes was similar. Vascular cells undergoing apoptosis were infrequent in both forms. CONCLUSION: Using four separate indices, apoptotic activity is more pronounced in cardiac allografts undergoing acute compared with chronic rejection. This reflects, in part, the degree of alloimmune response. However, we speculate that the contributions of apoptosis to various forms of rejection are multifactorial. The long-term outcome to the graft may depend upon the magnitude, timing, and target of programmed cell death.

Attenuated acute cardiac rejection in NOS2 -/- recipients correlates with reduced apoptosis.

BACKGROUND: The mechanisms through which NOS2-mediated pathways regulate graft failure in acute cardiac rejection are ill defined. To determine whether apoptosis promoted by NOS2 may contribute, we used a heterotopic transplant model to study mouse cardiac allografts placed in recipients with targeted gene deletion of NOS2. METHODS AND RESULTS: Using 5 different indexes of apoptosis, we showed that mouse cardiac allografts placed in NOS2 -/- recipients (n=7) had reduced apoptotic activity compared with those in NOS2 +/+ controls (n=8). There were significantly fewer TUNEL-positive nuclei per high-powered field (P<0.01), less DNA fragmentation (antinucleosome ELISA; P<0.05), lower corrected transcript levels for caspase-1 and -3 (32P reverse transcriptase-polymerase chain reaction; P<0.01), and reduced caspase-3 activity (cleavage of DEVD-pNA [P<0.001] and poly [ADP-ribose] polymerase) in grafts from NOS2 -/- recipients. This concordant reduction in apoptotic indexes paralleled the improved histological outcome of grafts transplanted into NOS2 -/- recipients (assessed as rejection scores; P=0.012). To identify pathways controlled by NOS2, we compared intragraft transcript levels of potential triggers and regulators. Whereas Fas ligand/Fas and tumor necrosis factor (TNF)-alpha/TNF receptor-1 levels were not altered by NOS2 deficiency, transcript levels for p53 were significantly lower in grafts from NOS2 -/- recipients, coinciding with a significant increase in the antiapoptotic Bcl-2/Bax balance and decrease in Bcl-Xl levels. CONCLUSIONS: Using NOS2 knockout mice, we demonstrated that NOS2-mediated pathways can promote acute rejection, at least in part, by inducing apoptotic cell death. When NOS2 is present, p53 might control NOS2-mediated apoptosis by stimulating Bax and repressing Bcl-2 and Bcl-Xl expression, which may activate the cell death program in the rejecting heart.

Mice with STAT6-targeted gene disruption develop a Th1 response and control cutaneous leishmaniasis.

The cutaneous growth of Leishmania mexicana was measured in STAT6-deficient mice (STAT6-/-) and compared with that in similarly infected wild-type (STAT6+/+) mice. Following s.c. inoculation with 5 x 10(6) amastigotes of L. mexicana into the shaven rump, STAT6+/+ mice developed large, nonhealing cutaneous lesions, while STAT6-/- mice failed to develop detectable lesions during most of the course of study. As infection progressed, STAT6+/+ mice infected with L. mexicana displayed significantly higher titers of Leishmania-specific IgG1 and IgE compared with STAT6-/- mice, which conversely produced significantly higher titers of Leishmania-specific IgG2a, indicating development of a Th1-like response in the latter group. At 12 wk postinfection, Leishmania Ag-stimulated lymph node cells from STAT6-/- mice produced significantly higher amounts of IL-12 and IFN-gamma than those from STAT6+/+ mice as measured by ELISA. However, there was no significant difference in IL-4 production between the two groups. Semiquantitative RT-PCR of transcript levels in intact draining lymph nodes and skin from inoculation sites confirmed a similar pattern of cytokines in vivo as that observed in stimulated lymph node cells in vitro. These results indicate that STAT6-mediated IL-4 signaling is critical for progression of L. mexicana infection in genetically susceptible mice and demonstrate that in the absence of STAT6, susceptible mice default toward a Th1-like response and control cutaneous L. mexicana infection.

Redefining peripheral tolerance in the BALB/c to CBA mouse cardiac allograft model: vascular and cytokine analysis after transient CD4 T cell depletion.

BACKGROUND: To evaluate cardiac allografts from recipients that had achieved peripheral tolerance after transient CD4+ T cell depletion, we analyzed cellular infiltrate, cytokine expression, and vascular thickening. Long-surviving cardiac allografts from tolerant recipients were compared with acutely rejecting allografts and isografts. METHODS AND RESULTS: In CBA mice treated with anti-CD4 (GK1.5, 0.5 mg intraperitoneally on days 1-28), BALB/c cardiac allografts survived >100 days. These recipients were tested for tolerance at >70 days, by challenge with donor and third-party (C57BL/6) skin grafts. BALB/c skin grafts survived >30 days, although C57BL/6 skin was rejected in <12 days, reflecting alloantigen-specific peripheral tolerance. When vascular thickening in graft arteries was assessed and computerized measurements performed, heart allografts from tolerant recipients showed significantly increased percentage of luminal occlusion compared with isografts (47% compared with 1.2%). Semiquantitative reverse transcriptase-polymerase chain reaction was used to assess normalized intragraft mRNA transcripts for cytokines and T cell markers, with immunoperoxidase staining of frozen sections to confirmed the presence of protein. Compared with rejecting grafts, well-preserved hearts from tolerant mice had lower levels of macrophage and T cell infiltration and decreased transcription of interferon-gamma, interleukin (IL)-2, IL-10, and inducible nitric oxide synthase. IL-4 expression was similar in both groups. CONCLUSIONS: The degree of tolerance achieved allowed specific acceptance of donor skin grafts, preserved primary graft function, and reduced inflammatory activation. Tolerance did not, however, completely prevent macrophage and T cell infiltration of the graft or the development of vascular lesions typical of chronic rejection.

NOS2 mediates opposing effects in models of acute and chronic cardiac rejection: insights from NOS2-knockout mice.

To compare regulatory effects of NOS2 in acute and chronic cardiac allograft rejection, we used NOS2 knockout mice as recipients in a cardiac transplant model. To study acute and chronic rejection separately but within the same genetic strain combination, we compared allografts placed into recipients without or with immunosuppression (anti-CD4/8 for 28 days). NOS2 mRNA and protein expression were compared using 32P-RT-PCR and immunohistochemistry. In our acute rejection model, NOS2 was predominately localized to graft-infiltrating immune cells. At day 7, grafts in NOS2-deficient recipients (n = 7) showed reduced inflammatory infiltrates and myocyte damage resulting in significantly lower rejection scores (1.6 +/- 0.4) compared to wild-type controls (n = 18; 2.8 +/- 0.2, P = 0.002). In contrast, in our chronic rejection model, additional NOS2 expression was localized to graft-parenchymal cells. At day 55, grafts in NOS2-deficient recipients (n = 12) showed more parenchymal infiltration and parenchymal destruction (rejection score 3.8 +/- 0.1) than wild-type controls (n = 15; 1.6 +/- 0.2, P < 0.0001). This was associated with a significant decrease in ventricular contractility (palpation score 0.3 +/- 0.1 compared to 2.3 +/- 0.3 in wild-type, P < 0.0001). Hence, NOS2 promotes acute but prevents chronic rejection. These opposing effects during acute and chronic cardiac allograft rejection are dependent on the temporal and spatial expression pattern of NOS2 during both forms of rejection.

Leukocyte-suppressing influences of interleukin (IL)-10 in cardiac allografts: insights from IL-10 knockout mice.

To investigate the role of interleukin (IL)-10 in late graft outcomes, we compared BALB/c donor hearts transplanted into immunosuppressed wild-type or IL-10 gene-deficient (-/-) C57BL recipients (n = 49) at 50 +/- 5 days. There was prominent leukocyte infiltration and parenchymal destruction with more severe vascular occlusion in grafts from IL-10 -/- recipients. An occlusive CD45+ arteritis with medial necrosis occurred with IL-10 deficiency instead of the a-smooth muscle actin-rich arteriosclerosis seen in wild-type recipients. Increased interferon (IFN)-gamma as well as Mac-1, inducible nitric oxide synthase, and allograft inflammatory factor-1 (but not CD3 and IL-4) transcript levels were seen in allografts from IL-10 -/- recipients as assessed by 32p reverse transcription polymerase chain reaction. We then evaluated the contribution of IFN-gamma-mediated responses by neutralizing IFN-gamma. Anti-IFN-gamma monoclonal antibody (MAb) treatment of IL-10 -/- recipients did not improve graft survival, parenchymal rejection, or occlusive arteritis, indicating that these processes are IFN-gamma independent. However, medial smooth muscle cell loss in IL-10 -/- recipients was attenuated by anti-IFN-gamma MAb. Hence, in this transplant model, IL-10 suppresses T cell and macrophage responses in the parenchyma and vasculature and confers a protective effect against late rejection.

Immune sources of transforming growth factor-beta1 reduce transplant arteriosclerosis: insight derived from a knockout mouse model.

Activated CD4-positive T cells are essential in the early stages of arteriosclerotic lesion development after cardiac transplantation. Besides its parenchymal effects, transforming growth factor-beta1 (TGF-beta1) mediates immunosuppressive effects on proliferation and activation of CD4 cells. This study was designed to assess immune contributions of TGF-beta1 to arteriosclerosis by comparing the effect of TGF-beta1-deficient and -competent infiltrating inflammatory cells on the development of intimal thickening in a heterotopic mouse transplant model (CBA to C57B6). Transplant arteriosclerosis was evaluated in cardiac grafts placed into knockout recipients heterozygous for TGF-beta1 (n=7) and was compared with those placed into wild-type recipients (n=11). At 55 days, allografts in TGF-beta1-deficient recipients had increased concentric intimal thickening. Computer-assisted analysis of all elastin-positive vessels (n=173) showed significantly increased luminal occlusion (67.8+/-5.6%) in grafts from TGF-beta1-deficient recipients compared with wild-type recipients (47.4+/-4.1%, P=0.003). To determine whether TGF-beta1 deficiency altered CD4 activation patterns, we studied intragraft cytokine expression. Using 32P-reverse-transcriptase polymerase chain reaction assays, we show that TGF-beta1-deficient recipients had an increased expression of the transcription factor STAT 4, interferon gamma, and interleukin-2 (Th1-type response) and unaltered or reduced expression of the transcription factor STAT 6, interleukin-4, and interleukin-10 (Th2-type response). Hence, when present, immune sources of TGF-beta1 attenuate transplant arteriosclerosis. This effect is associated with attenuation of Th1 forces.

Cardiac allografts from IL-4 knockout recipients: assessment of transplant arteriosclerosis and peripheral tolerance.

To study the role of IL-4 in tolerance induction and transplant arteriosclerosis, BALB/c hearts were transplanted into C57BL/6J wild-type or IL-4 knockout (IL-4(-/-)) recipients. A 30-day course of anti-CD4/8 mAb was used to induce long term graft survival. Primary graft survival was 50% (5 of 10) in IL-4(-/-) recipients comparable to 63% (5 of 8) in wild-type recipients. Mice with allografts surviving >80 days were tested for tolerance by challenge with a second donor or third party (CBA) heart. Secondary donor-strain heart grafts survived >30 days, but showed histologic evidence of ongoing alloimmune response. Third party hearts rejected rapidly. Although immunostaining and 32P RT-PCR assays showed no differences in the mononuclear cell infiltration and T cell activation between IL-4(-/-) and wild-type tolerant recipients, some monokines (IL-12, TNF-alpha, and allograft inflammatory factor-1) were up-regulated in grafts from IL-4(-/-) recipients. Computer-assisted analysis of elastin-stained vessels revealed that the severity of vascular thickening (percentage of luminal occlusion, mean +/- SD, n = 329) was similar in grafts from IL-4(-/-) (63.7 +/- 16.9%) and wild-type (69.5 +/- 17.6%) recipients. Thus, IL-4 deficiency did not alter primary or secondary graft survival, infiltration, or vascular thickening. The selective alterations in monokine expression suggests that alternative pathways are activated and may compensate in IL-4(-/-) mice.