Chloroquine (CQ) resistance in the human malaria parasite Plasmodium falciparum is primarily conferred by mutations in the “chloroquine resistance transporter” (PfCRT). The resistance-conferring form of PfCRT (PfCRTCQR) mediates CQ resistance by effluxing the drug from the parasite’s digestive vacuole, the acidic compartment in which CQ exerts its antiplasmodial effect. PfCRTCQR can also decrease the parasite’s susceptibility to other quinoline drugs, including the current antimalarials quinine and amodiaquine. Here we describe interactions between PfCRTCQR and a series of dimeric quinine molecules using a Xenopus laevis oocyte system for the heterologous expression of PfCRT and using an assay that detects the drug-associated efflux of H+ ions from the digestive vacuole in parasites that harbor different forms of PfCRT. The antiplasmodial activities of dimers 1 and 6 were also examined in vitro (against drug-sensitive and drug-resistant strains of P. falciparum) and in vivo (against drug-sensitive P. berghei). Our data reveal that the quinine dimers are the most potent inhibitors of PfCRTCQR reported to date. Furthermore, the lead compounds (1 and 6) were not effluxed by PfCRTCQR from the digestive vacuole but instead accumulated to very high levels within this organelle. Both 1 and 6 exhibited in vitro antiplasmodial activities that were inversely correlated with CQ. Moreover, the additional parasiticidal effect exerted by 1 and 6 in the drug-resistant parasites was attributable, at least in part, to their ability to inhibit PfCRTCQR. This highlights the potential for devising new antimalarial therapies that exploit inherent weaknesses in a key resistance mechanism of P. falciparum.
Clinical data have shown that tick-borne diseases caused by Borrelia burgdorferi sensu lato, Bartonella spp., Coxiella burnetii, and Rickettsia spp. can affect the central nervous system, including the eye. The aim of this study was to establish a relationship between the incidence of cataract and evidence of bacterial infections transmitted by ticks.
Fluid with lenticular masses from inside of the eye and blood from 109 patients were tested by PCR and sequencing. Sera from patients and the control group were subjected to serological tests to search specific antibodies to the bacteria.
Microbiological analysis revealed the presence of Bartonella sp. DNA in intraoperative specimens from the eye in 1.8% of patients. Serological studies have shown that infections caused by B. burgdorferi sensu lato and Bartonella sp. were detected in 34.8% and 4.6% of patients with cataract surgery, respectively.
Presence of DNA of yet uncultured and undescribed species of Bartonella in eye liquid indicates past infection with this pathogen. Specific antibodies to B. burgdorferi sensu lato and Bartonella sp. are detected more frequently in patients with cataract compared to the control group. This could indicate a possible role of these organisms in the pathological processes within the eyeball, leading to changes in the lens. Further studies are needed to identify Bartonella species, as well as to recognize the infectious mechanisms involved in cataract development.
Borrelia Burgdorferi Sensu Lato; Bartonella Spp.; Coxiella Burnetii; Rickettsia Spp.; Cataract
Smokers are more prone to oral infections than are non-smokers. Cigarette smoke reaches the host cells but also microorganisms present in the oral cavity. The contact between cigarette smoke and oral bacteria promotes such oral diseases as periodontitis. Cigarette smoke can also modulate C. albicans activities that promote oral candidiasis. The goal of this study was to investigate the effect of cigarette smoke condensate on C. albicans adhesion, growth, and biofilm formation as well as the activation of EAP1, HWP1 and secreted aspartic protease 2.
Cigarette smoke condensate (CSC) increased C. albicans adhesion and growth, as well as biofilm formation. These features may be supported by the activation of certain important genes. Using quantitative RT-PCR, we demonstrated that CSC-exposed C. albicans expressed high levels of EAP1, HWP1 and SAP2 mRNA and that this gene expression increased with increasing concentrations of CSC.
CSC induction of C. albicans adhesion, growth, and biofilm formation may explain the increased persistence of this pathogen in smokers. These findings may also be relevant to other biofilm-induced oral diseases.
Cigarette smoke; Tobacco; C albicans; Adhesion; Growth; Biofilm; Genes; EAP-1; HWP-1; Sap2
Metabolic syndrome (MetS) is associated with increased risk of cardiovascular disease (CVD). One important feature underlying the pathophysiology of many types of CVD is microvascular dysfunction. Although components of MetS are themselves CVD risk factors, the risk is increased when the syndrome is considered as one entity. We aimed to characterize microvascular function and some of its influencing factors in the course of MetS development.
Development of MetS in C57BL/6 mice on a high-fat diet (HFD, 51% of energy from fat) was studied. The initial phase of MetS (I-MetS) was defined as the first 2 weeks of HFD feeding, with the fully developed phase occurring after 8 weeks of HFD. We characterized these phases by assessing changes in adiposity, blood pressure, and microvascular function. All data are presented as mean ± standard error (SEM). Differences between cumulative dose–response curves of myograph experiments were calculated using non-linear regression analysis. In other experiments, comparisons between two groups were made with Student’s t-test. Comparisons between more than two groups were made using one-way ANOVA with Tukey post-hoc test. A probability value <0.05 was considered statistically significant.
I-MetS mice presented with weight gain, blood pressure elevation, and microvascular dysfunction characterized by augmented vasoconstriction. This finding, contrary to those in mice with fully developed MetS, was not associated with endothelial dysfunction, insulin resistance, or systemic inflammation. In the initial phase, perivascular adipose tissue showed no sign of inflammation and had no influence on the pattern of vasoconstriction. These findings suggest that the onset of hypertension in MetS is strongly influenced by vascular smooth muscle cell dysfunction and independent of important factors known to influence microvascular function and consequently blood pressure levels.
We identified in I-MetS the occurrence of isolated augmented vasoconstriction along with blood pressure elevation, but not the presence of classical MetS components known to influence microvascular function. These findings increase our understanding of the pathophysiology of CVD risk associated with MetS.
High-fat diet; Metabolic syndrome; Hypertension; Microvascular dysfunction; Vasoconstriction
Post-traumatic knee osteoarthritis is prevalent after anterior cruciate ligament reconstruction. Biomarkers that identify individuals likely to develop osteoarthritis, especially symptomatic osteoarthritis, can help target preventative and therapeutic strategies. This study examined the magnitude and change over time in urinary CTX-II (uCTX-II) concentrations shortly after ACL reconstruction, and, secondarily, the associations with knee pain and function.
Subjects were 28 patients with ACL reconstruction (ACLR) and 28 age- and sex-matched controls (CNTRL). Testing was conducted at 4 time points spaced 4 weeks apart (4, 8, 12 and 16 weeks post-operative in ACLR). Measures included demographics, urine samples, Numeric Pain Rating Scale (NPRS) and International Knee Documentation Committee Subjective Knee Form (IKDC-SKF). uCTX-II concentrations were determined with competitive ELISA. uCTX-II concentrations at each time point in ACLR were compared to the mean concentration over time in CNTRL, with and without adjustment for body mass index (BMI). Changes over time in each measure and correlations between the slopes of change were examined.
uCTX-II concentrations were significantly higher in ACLR than CNTRL through 16 weeks post-operative when adjusted for BMI. In ACLR, uCTX-II concentrations significantly decreased over time, and the slope was associated with NPRS (r =.406, p=.039) and IKDC-SKF (r = −.402, p = .034) slopes.
uCTX-II concentrations shortly after ACL reconstruction were elevated compared to controls and declined over time. Decreasing uCTX-II concentrations were associated with decreasing knee pain and improving function. uCTX-II may have a role as a prognostic marker following ACL reconstruction and warrants further investigation.
Maternal parenting self-efficacy (PSE) is a potential target for infant mental health interventions because it is associated with a number of positive outcomes for children and mothers. Understanding the development of maternal PSE under conditions of increased parenting stress, such as parenting an infant who is easily distressed and difficult to soothe, will contribute to providing more effective interventions. This study examines the development of maternal PSE in mothers of infants with high negative emotionality (NE). The Neonatal Behavioral Assessment Scale (NBAS; T. Brazelton, 1973) was administered twice to 111 infants to select a sample of irritable (n = 24) and nonirritable (n = 29) infants for a prospective study comparing the development of PSE in mothers of infants differing in neonatal NE. Consistent with our hypotheses and previous research, at 8 weeks’ postpartum, mothers of irritable infants have significantly lower domain-specific PSE than do mothers of nonirritable infants. Contrary to our predictions, mothers of irritable infants exhibit a significant increase in domain-specific and domain-general PSE from 8 to 16 weeks’ postpartum. The implications of these results for infant mental health screening, infant mental health interventions, and research on self-efficacy theory are discussed.
The condensation state of chromosomes is a critical parameter in multiple processes within the cell. Failures in the maintenance of appropriate condensation states may lead to genomic instability, mis-expression of genes, and a number of disease states. During cell proliferation, replication of DNA represents an ongoing challenge for chromosome packaging as DNA must be unpackaged for replication and then faithfully repackaged. An integral member of the DNA replication machinery is the GINS complex which has been shown to stabilize the CMG complex which is required for processivity of the Mcm2–7 helicase complex during S phase. Through examination of the phenotypes associated with a null mutation in Psf2, a member of the evolutionarily conserved GINS complex, we find that Drosophila Psf2 likely has a role in establishing chromosome condensation and that the defects associated with this mis-condensation impact M phase progression, genomic stability, and transcriptional regulation.
GINS Complex; Psf2; DNA replication; Chromosome condensation
Obsessive-compulsive disorder (OCD) is a heterogeneous condition, comprised of multiple symptom domains. This study used aggregate composite scales representing three core OCD dimensions (Checking, Cleaning, Rituals), as well as Hoarding, to examine the discriminant validity, diagnostic specificity, and predictive ability of OCD symptom scales. The core OCD scales demonstrated strong patterns of convergent and discriminant validity – suggesting that these dimensions are distinct from other self-reported symptoms – whereas hoarding symptoms correlated just as strongly with OCD and non-OCD symptoms in most analyses. Across analyses, our results indicated that Checking is a particularly strong, specific marker of OCD diagnosis, whereas the specificity of Cleaning and Hoarding to OCD was less strong. Finally, the OCD Checking scale was the only significant predictor of OCD diagnosis in logistic regression analyses. Results are discussed with regard to the importance of assessing OCD symptom dimensions separately and implications for classification.
obsessive compulsive disorder; symptoms; specificity; hoarding; clinical population; discriminant validity
A confirmed case of rickettsiosis acquired in South Africa and recognized in Poland was described. The patient fulfilled clinical criteria highly suggestive of African tick bite fever, such as eschars, regional lymphadenitis, cutaneous rash within 10 days after his return from sub-Saharan Africa. Infection with Rickettsia africae was confirmed by polymerase chain reaction and sequencing.
Rickettsia africae; polymerase chain reaction sequencing; imported infection
Q fever is a health problem affecting humans and animals worldwide. In Poland, previous studies have pointed to 2 sources of outbreaks of the disease: the importation of infected animals and their products, and natural domestic foci. In the last decade, 5 outbreaks have occurred in cattle farms in south Poland in Malopolskie, Podkarpackie, Opolskie, and Silesian provinces. The aim of this study was to characterize the Q fever foci in Poland.
A total of 279 individuals were included. Levels of serum IgM and IgG antibodies to phase I and II C. burnetii antigens were assayed by indirect immunofluorescence method. Bacterial DNA from all specimens were detected with PCR with primer pairs specific to the htpAB-associated repetitive element, and amplicons were sequenced.
Infection was recognized in 67 individuals out of 279 tested in all foci. Twenty-five individuals presented clinical symptoms of acute Q fever. DNA of C. burnetii was found in 8 human blood samples obtained from 3 farm workers and 5 family members.
The described outbreaks demonstrate that the main source of human infections in Poland is infected cattle.
Coxiella burnetii; Q fever; outbreaks
Hepatitis C virus (HCV) coinfection was reported to negatively affect HIV disease and HIV infection has a deleterious effect on HCV-related liver disease. However, despite common occurrence of HCV/HIV coinfection little is known about the mechanisms of interactions between the two viruses.
We studied CD4+ and CD8+ T cell and CD19+ B cell apoptosis in 104 HIV-positive patients (56 were also HCV-positive) and in 22 HCV/HIV-coinfected patients treated for chronic hepatitis C with pegylated interferon and ribavirin. We also analyzed HCV/HIV coinfection in a Daudi B-cell line expressing CD4 and susceptible to both HCV and HIV infection. Apoptosis was measured by AnnexinV staining.
HCV/HIV coinfected patients had lower CD4+ and CD8+ T cell apoptosis and higher CD19+ B cell apoptosis than those with HIV monoinfection. Furthermore, anti-HCV treatment of HCV/HIV coinfected patients was followed by an increase of CD4+ and CD8+ T cell apoptosis and a decrease of CD19+ B cell apoptosis. In the Daudi CD4+ cell line, presence of HCV infection facilitated HIV replication, however, decreased the rate of HIV-related cell death.
In HCV/HIV coinfected patients T-cells were found to be destroyed at a slower rate than in HIV monoinfected patients. These results suggest that HCV is a molecular-level determinant in HIV disease.
Recent work suggests that psychological influence on pain intensity and knee function should be considered for patients following anterior cruciate ligament reconstruction (ACLR). The Tampa Scale for Kinesiophobia (TSK) and Pain Catastrophizing Scale (PCS) have been used to determine psychological influence in these patients. However, TSK and PCS factor structures have not been described for patients with ACLR. This study investigated 2 groups of patients post-ACLR to determine if the use of shortened questionnaires is warranted.
This was a cross-sectional study in which patients completed measures during early (n = 105, median days from surgery = 56.0) and late (n = 184, median days from surgery = 195.0) post-operative phases of ACLR rehabilitation.
Shortened questionnaires for fear of pain, fear of injury, and somatic focus were generated for the TSK-11. A shortened questionnaire for magnification/helplessness and rumination were generated for the PCS in the late group only. There were minimal differences in the shortened questionnaires for clinical subgroups based on sex, ACLR graft type, method of injury, or nature of injury. Correlation and regression analyses suggested a shortened version of the TSK-11 for fear of injury was appropriate for use in the early post-operative phase, while the original TSK-11 scale may be appropriate for use in the late post-operative phase. There were no shortened versions of the PCS for consideration in the early post-operative phase, but a shortened version for magnification/helplessness was appropriate for use in the late post-operative phase.
Shortened versions of the TSK-11 and PCS may be appropriate for ACLR populations, depending on the post-operative phase. These data may guide future research of psychological factors in ACLR populations so that levels predictive of risk for developing chronic pain and/or inability to return to pre-injury activity levels can be determined.
Chronic pain; pain catastrophizing; fear-avoidance; knee function; rehabilitation
(1) To examine differences in clinical variables (demographics, knee impairments, and self-report measures) between those who return to preinjury level of sports participation and those who do not at 1 year following anterior cruciate ligament reconstruction, (2) to determine the factors most strongly associated with return-to-sport status in a multivariate model, and (3) to explore the discriminatory value of clinical variables associated with return to sport at 1 year postsurgery.
Demographic, physical impairment, and psychosocial factors individually prohibit return to preinjury levels of sports participation. However, it is unknown which combination of factors contributes to sports participation status.
Ninety-four patients (60 men; mean age, 22.4 years) 1 year post–anterior cruciate ligament reconstruction were included. Clinical variables were collected and included demographics, knee impairment measures, and self-report questionnaire responses. Patients were divided into “yes return to sports” or “no return to sports” groups based on their answer to the question, “Have you returned to the same level of sports as before your injury?” Group differences in demographics, knee impairments, and self-report questionnaire responses were analyzed. Discriminant function analysis determined the strongest predictors of group classification. Receiver-operating-characteristic curves determined the discriminatory accuracy of the identified clinical variables.
Fifty-two of 94 patients (55%) reported yes return to sports. Patients reporting return to preinjury levels of sports participation were more likely to have had less knee joint effusion, fewer episodes of knee instability, lower knee pain intensity, higher quadriceps peak torque-body weight ratio, higher score on the International Knee Documentation Committee Subjective Knee Evaluation Form, and lower levels of kinesiophobia. Knee joint effusion, episodes of knee instability, and score on the International Knee Documentation Committee Subjective Knee Evaluation Form were identified as the factors most strongly associated with self-reported return-to-sport status. The highest positive likelihood ratio for the yes-return-to-sports group classification (14.54) was achieved when patients met all of the following criteria: no knee effusion, no episodes of instability, and International Knee Documentation Committee Subjective Knee Evaluation Form score greater than 93.
In multivariate analysis, the factors most strongly associated with return-to-sport status included only self-reported knee function, episodes of knee instability, and knee joint effusion.
ACL; kinesiophobia; return to sports
Chronic hepatitis C viremia (HepC) has been associated with numerous renal manifestations both in native kidneys and in the setting of renal transplantation. Glomerulonephritis (GN) of the renal allograft in the setting of HepC most commonly manifests as type 1 membranoproliferative GN (MPGN), either representing recurrence of the original disease or arising de novo. Other GNs were reported after transplantation in the patient with HepC including membranous nephropathy and thrombotic microangiopathy, as well as an enhanced susceptibility to transplant glomerulopathy. We describe the first case of de novo fibrillary GN in a renal transplant patient with HepC where the primary renal disease was biopsy proven type 1 MPGN. We discuss this relationship in detail.
In response to the Association of American Medical Colleges–Howard Hughes Medical Institute report Scientific Foundations for Future Physicians, a collaborative effort by four institutions has produced an introductory physics for life sciences course that stresses competency building and helps students apply strategies from the physical sciences to solve authentic biological problems.
The National Experiment in Undergraduate Science Education project funded by the Howard Hughes Medical Institute is a direct response to the Scientific Foundations for Future Physicians report, which urged a shift in premedical student preparation from a narrow list of specific course work to a more flexible curriculum that helps students develop broad scientific competencies. A consortium of four universities is working to create, pilot, and assess modular, competency-based curricular units that require students to use higher-order cognitive skills and reason across traditional disciplinary boundaries. Purdue University; the University of Maryland, Baltimore County; and the University of Miami are each developing modules and case studies that integrate the biological, chemical, physical, and mathematical sciences. The University of Maryland, College Park, is leading the effort to create an introductory physics for life sciences course that is reformed in both content and pedagogy. This course has prerequisites of biology, chemistry, and calculus, allowing students to apply strategies from the physical sciences to solving authentic biological problems. A comprehensive assessment plan is examining students’ conceptual knowledge of physics, their attitudes toward interdisciplinary approaches, and the development of specific scientific competencies. Teaching modules developed during this initial phase will be tested on multiple partner campuses in preparation for eventual broad dissemination.
To determine the association of two dimensional (2D) video-based techniques and three-dimensional (3D) motion analysis to assess potential knee injury risk factors during jump landing.
Thirty-six female athletes in cutting and pivoting sports.
Athletes performed a drop vertical jump during which movement was recorded with a motion analysis system and a digital video camera positioned in the frontal plane.
Main Outcome Measures
The 2D variables were the frontal plane projection angle (FPPA), the angle formed between thigh and leg, and the knee:ankle separation ratio, the distance between knee joints divided by the distance between ankles. The 3D variables were knee abduction angle and external abduction moment. All variables were assessed at peak knee flexion. Linear regression assessed the relationship between the 2D and 3D variables. In addition, intraclass correlation coefficients determined rater reliability for the 2D variables and compared the 2D measurements made from digital video to the same measurements from the motion analysis.
The knee:ankle separation ratio accounted for a higher variance of 3D knee abduction angle (r2 =0.350) and knee abduction moment (r2=0.394) when compared to the FPPA (r2=0.145, 0.254). The digital video measures had favorable rater reliability (ICC:0.89–0.94) and were comparable to the motion analysis system (ICC≥0.92).
When compared to the FPPA, the knee:ankle separation ratio had better association with previously cited knee injury risk factors in female athletes. The 2D measures have adequate consistency and validity to merit further clinical consideration in jump landing assessments.
ACL injury prevention; assessment tools; female athletes
Variations in joint parameter values (axis positions and orientations in body segments) and inertial parameter values (segment masses, mass centers, and moments of inertia) as well as kinematic noise alter the results of inverse dynamics analyses of gait. Three-dimensional linkage models with joint constraints have been proposed as one way to minimize the effects of noisy kinematic data. Such models can also be used to perform gait optimizations to predict post-treatment function given pre-treatment gait data. This study evaluates whether accurate patient-specific joint and inertial parameter values are needed in three-dimensional linkage models to produce accurate inverse dynamics results for gait. The study was performed in two stages. First, we used optimization analyses to evaluate whether patient-specific joint and inertial parameter values can be calibrated accurately from noisy kinematic data, and second, we used Monte Carlo analyses to evaluate how errors in joint and inertial parameter values affect inverse dynamics calculations. Both stages were performed using a dynamic, 27 degree-of-freedom, full-body linkage model and synthetic (i.e., computer generated) gait data corresponding to a nominal experimental gait motion. In general, joint but not inertial parameter values could be found accurately from noisy kinematic data. Root-mean-square (RMS) errors were 3° and 4 mm for joint parameter values and 1 kg, 22 mm, and 74,500 kg*mm2 for inertial parameter values. Furthermore, errors in joint but not inertial parameter values had a significant effect on calculated lower-extremity inverse dynamics joint torques. The worst RMS torque error averaged 4% bodyweight*height (BW*H) due to joint parameter variations but less than 0.25% BW*H due to inertial parameter variations. These results suggest that inverse dynamics analyses of gait utilizing linkage models with joint constraints should calibrate the model’s joint parameter values to obtain accurate joint torques.
joint parameters; body segment parameters; inverse dynamics; linkage models; gait
Differences in various outcome measures have been identified between people who have sprained their ankles but have no residual symptoms (copers) and people with chronic ankle instability (CAI). However, the diagnostic utility of the reported outcome measures has rarely been determined. Identifying outcome measures capable of predicting who is less likely to develop CAI could improve rehabilitation protocols and increase the efficiency of these measures.
To determine the diagnostic utility and cutoff scores of perceptual, mechanical, and sensorimotor outcome measures between copers and people with CAI by using receiver operating characteristic curves.
Sports medicine research laboratory.
Patients or Other Participants:
Twenty-four copers (12 men, 12 women; age = 20.8 ± 1.5 years, height = 173 ± 11 cm, mass = 78 ± 27 kg) and 24 people with CAI (12 men, 12 women; age = 21.7 ± 2.8 years, height = 175 ± 13 cm, mass = 71 ± 13 kg) participated.
Self-reported disability questionnaires, radiographic images, and a single-legged hop stabilization test.
Main Outcome Measure(s):
Perceptual outcomes included scores on the Foot and Ankle Disability Index (FADI), FADI-Sport, and a self-report questionnaire of ankle function. Mechanically, talar position was quantified by measuring the distance from the anterior tibia to the anterior talus in the sagittal plane. Sensorimotor outcomes were the dynamic postural stability index and directional indices, which were calculated during a single-legged hop stabilization task.
Perceptual outcomes demonstrated diagnostic accuracy (range, 0.79–0.91), with 95% confidence intervals ranging from 0.65 to 1.00. Sensorimotor outcomes also were able to discriminate between copers and people with CAI but with less accuracy (range, 0.69–0.70), with 95% confidence intervals ranging from 0.37 to 0.86. The mechanical outcome demonstrated poor diagnostic accuracy (0.52).
The greatest diagnostic utility scores were achieved by the self-assessed disability questionnaires, which indicated that perceptual outcomes had the greatest ability to accurately predict people who became copers after their initial injuries. However, the diversity of outcome measures that discriminated between copers and people with CAI indicated that the causal mechanism of CAI is probably multifactorial.
self-report disability; positional fault; dynamic postural control
Eradication of HIV reservoirs in the brain necessitates penetration of antiviral agents across the blood-brain barrier (BBB), a process limited by drug efflux proteins such as P-glycoprotein (P-gp) at the membrane of brain capillary endothelial cells. We present an innovative chemical strategy toward the goal of therapeutic brain penetration of the P-gp substrate and anti-viral agent abacavir, in conjunction with a traceless tether. Dimeric prodrugs of abacavir were designed to have two functions: inhibit P-gp efflux at the BBB and revert to monomeric therapeutic within cellular reducing environments. The prodrug dimers are potent P-gp inhibitors in cell culture and in a brain capillary model of the BBB. Significantly, these agents demonstrate anti-HIV activity in two T-cell-based HIV assays, a result that is linked to cellular reversion of the prodrug to abacavir. This strategy represents a platform technology that may be applied to other therapies with limited brain penetration due to P-glycoprotein.
Adoptive T-cell therapy has recently shown promise in initiating a lasting anti-tumor response with spectacular therapeutic success in some cases. Specific T-cell therapy, however, is limited since a number of cancer cells are not recognized by T cells due to various mechanisms including the limited availability of tumor-specific T cells and deficiencies in antigen processing or major histocompatibility complex (MHC) expression of cancer cells. To make adoptive cell therapy applicable for the broad variety of cancer entities, patient’s T cells are engineered ex vivo with pre-defined specificity by a recombinant chimeric antigen receptor (CAR) which consists in the extracellular part of an antibody-derived domain for binding with a “tumor-associated antigen” and in the intracellular part of a T-cell receptor (TCR)-derived signaling moiety for T-cell activation. The specificity of CAR-mediated T-cell recognition is defined by the antibody domain, is independent of MHC presentation and can be extended to any target for which an antibody is available. We discuss the advantages and limitations of MHC-independent T-cell targeting by an engineered CAR in comparison to TCR modified T cells and the impact of the CAR activation threshold on redirected T-cell activation. Finally we review most significant progress recently made in early stage clinical trials to treat cancer.
chimeric antigen receptor; T-cell receptor; adoptive cell therapy; antibody; antigen-presenting cell
The gingival epithelium is becoming known as a regulator of the oral innate immune responses to a variety of insults such as bacteria and chemicals, including those chemicals found in cigarette smoke. We investigated the effects of whole cigarette smoke on cell-surface-expressed Toll-like receptors (TLR)-2, −4 and −6, human β-defensin (HBD) and proinflammatory cytokine expression and production in primary human gingival epithelial cells. Whole cigarette smoke was shown to increase TLR2, TLR4 and TLR6 expression. Cigarette smoke led to ERK1/2, p38 and JNK phosphorylation in conjunction with nuclear factor-κB (NFκB) translocation into the nucleus. TLR expression following cigarette smoke exposure was down regulated by the use of ERK1/2, p38, JNK MAP kinases, and NFκB inhibitors, suggesting the involvement of these signaling pathways in the cellular response against cigarette smoke. Cigarette smoke also promoted HBD2, HBD3, IL-1β, and IL-6 expression through the ERK1/2 and NFκB pathways. Interestingly, the modulation of TLR, HBD, and cytokine expression was maintained long after the gingival epithelial cells were exposed to smoke. By promoting TLR, HBDs, and proinflammatory cytokine expression and production, cigarette smoke may contribute to innate immunity dysregulation, which may have a negative effect on human health.
Despite intensive treatment, pancreatic adenocarcinoma still has the worst prognosis among all malignancies. Using clinically relevant models, we demonstrated the therapeutic efficacy of adoptively transferred T cells engineered with a carcinoembryonic antigen (CEA)- and ERBB2-specific chimeric antigen receptor against pancreatic carcinoma. Targeting CD24, a putative cancer stem cell antigen expressed by a minority of carcinoma cells, was likewise effective.
adoptive cell therapy; cancer stem cell; chimeric antigen receptor; pancreas adenocarcinoma; T cell
After peripheral nerve injury, neurotrophins play a key role in the regeneration of damaged axons which can be augmented by exercise, although the distinct roles played by neurons and Schwann cells are unclear. In this study, we evaluated the requirement for the neurotrophin, brain derived neurotrophic factor (BDNF), in neurons and Schwann cells, for the regeneration of peripheral axons after injury. Common fibular or tibial nerves in thy-1-YFP-H mice were cut bilaterally and repaired using a graft of the same nerve from transgenic mice lacking BDNF in Schwann cells (BDNF-/-) or wild-type mice (WT). Two weeks post-repair, axonal regeneration into BDNF-/- grafts were markedly less than WT grafts, emphasizing the importance of Schwann cell BDNF. Nerve regeneration was enhanced by treadmill training post-transection, regardless of the BDNF content of the nerve graft. We further tested the hypothesis that training-induced increases in BDNF in neurons allow regenerating axons to overcome a lack of BDNF expression in cells in the pathway through which they regenerate. Nerves in mice lacking BDNF in YFP+ neurons (SLICK) were cut and repaired with BDNF-/- and WT nerves. SLICK axons lacking BDNF did not regenerate into grafts lacking Schwann cell BDNF. Treadmill training could not rescue the regeneration into BDNF-/- grafts if the neurons also lacked BDNF. Both Schwann cell- and neuron-derived BDNF are thus important for axon regeneration in cut peripheral nerves.
Atherosclerosis is characterized by early endothelial dysfunction and altered vascular smooth muscle cells (VSMCs) contractility. The forming atheroma is a site of excessive production of cytokines and inflammatory ligands by various cell types that mediate inflammation and immune responses. Key factors contributing to early stages of plaque development are IFNγ and TLR4. This review provides insight in the differential STAT1-dependent signal integration between IFNγ and TLR4 signals in vascular cells and atheroma interacting immune cells. This results in increased leukocyte attraction and adhesion and VSMC proliferation and migration, which are important characteristics of EC dysfunction and early triggers of atherosclerosis.
atherosclerosis; IFN-gamma; Toll-like receptors; STAT1; IRFs; signal integration
Adoptive therapy with chimeric antigen receptor (CAR) redirected T cells recently showed remarkable anti-tumor efficacy in early phase clinical trials; self-repression of the immune response by T-cell secreted cytokines, however, is still an issue raising interest to abrogate the secretion of repressive cytokines while preserving the panel of CAR induced pro-inflammatory cytokines. We here revealed that T-cell activation by a CD28-ζ signaling CAR induced IL-10 secretion, which compromises T cell based immunity, along with the release of pro-inflammatory IFN-γ and IL-2. T cells stimulated by a ζ CAR without costimulation did not secrete IL-2 or IL-10; the latter, however, could be induced by supplementation with IL-2. Abrogation of CD28-ζ CAR induced IL-2 release by CD28 mutation did not reduce IL-10 secretion indicating that IL-10 can be induced by both a CD28 and an IL-2 mediated pathway. In contrast to the CD28-ζ CAR, a CAR with OX40 (CD134) costimulation did not induce IL-10. OX40 cosignaling by a 3rd generation CD28-ζ-OX40 CAR repressed CD28 induced IL-10 secretion but did not affect the secretion of pro-inflammatory cytokines, T-cell amplification or T-cell mediated cytolysis. IL-2 induced IL-10 was also repressed by OX40 co-signaling. OX40 moreover repressed IL-10 secretion by regulatory T cells which are strong IL-10 producers upon activation. Taken together OX40 cosignaling in CAR redirected T cell activation effectively represses IL-10 secretion which contributes to counteract self-repression and provides a rationale to explore OX40 co-signaling CARs in order to prolong a redirected T cell response.
CD28; IL-10; OX40; adoptive cell therapy; chimeric antigen receptor