Next generation antibody and TCR therapeutics for infectious disease

Parasitic and infectious diseases cause over 9.5 million deaths worldwide annually, yet only 3 of the over 50 approved antibodies are for infectious conditions. This disparity can be attributed to the high costs of antibody development in the face of small molecule alternatives and effective vaccines; however, a growing niche of specialty applications and the emergence of antibioticresistant strains make antibody therapeutics a likely eventuality. Particular challenges for developing therapeutic proteins for pathogenic diseases are that high variability in circulating strains, mutability within the host, and immune escape mechanisms limits the efficacy of monoclonal antibody formats. Instead, next generation formats that can exhibit broader activity or induce novel immune mechanisms may be a viable approach. T-cell receptors (TCRs) are membrane-bound molecules that bind peptide-MHC (pMHC) molecules displayed by host cells, and are experts at recognizing infected cells. We have replaced one arm of a bispecific antibody with a soluble TCR to make a novel TCR/Ig hybrid. To do this, we first developed a system which can display TCRs on the surface of CHO cells. After optimization of the TCR format, in particular constant region modifications, we replaced the transmembrane region of the construct with an IgG1 hinge and Fc to express and purify soluble fusions. By including a knob or hole mutation in the Fc region, we were able to make bispecific molecules. Our current format couples an anti-CD3 antibody specificity to a human TCR that is associated with cytomegalovirus (CMV). We hypothesize that this format may have unique ability to suppress CMV reactivation or infection in transplant recipients or pregnant women. Ongoing work includes affinity maturation of the TCR, as well as testing the bispecific in a cellular model of CMV

Retrospective evaluation of antimicrobial prophylaxis in prevention of surgical site infection in the pediatric population

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/108107/1/pan12436.pd

Perspectives of Women in Prostitution diversion program on DNA Collection for a High-Risk DNA Database

DNA can be collected from women at high risk of violent crime as a pre-emptive biometric for post-mortem identification. We conducted focus groups with women in a prostitution diversion program that offers pre-emptive DNA collection. In general, the women supported the program but voiced concern for law enforcement collecting DNA. These data provide insights into the challenges of collecting DNA from women whom law enforcement might consider alternately to be victims or vilified. Hearing the voices of these women provides the forensic community an opportunity to design programs to minimize harm and maximize utility of DNA for victim identification

Pneumococcal vaccination coverage among children with sickle cell anemia, sickle cell trait, and normal hemoglobin

BackgroundChildren with sickle cell anemia and sickle cell trait are at an increased risk of invasive pneumococcal disease compared to children with normal hemoglobin. We assessed and compared pneumococcal vaccination status among these three groups.ProcedureChildren with sickle cell anemia and sickle cell trait were identified using Michigan newborn screening records (1997–2014); each child was matched to four children with normal hemoglobin based on age, Medicaid enrollment (at least 1 year from 2012–2014), race, and census tract. Vaccination records were obtained from the state’s immunization system. Pneumococcal vaccine coverage (PCV7 or PCV13 depending on date of administration) was assessed at milestone ages of 3, 5, 7, and 16 months. The proportion of children with vaccine coverage at each milestone was calculated overall and compared among children with sickle cell anemia, sickle cell trait, and normal hemoglobin using chi‐square tests.ResultsThe study population consisted of 355 children with sickle cell anemia, 17,319 with sickle cell trait, and 70,757 with normal hemoglobin. The proportion of children with age‐appropriate pneumococcal vaccination coverage was low at each milestone and generally decreased over time. Children with sickle cell anemia were more likely to be covered compared to children with sickle cell trait or normal hemoglobin.ConclusionsDespite higher pneumococcal vaccination coverage among children with sickle cell anemia, opportunities for improvement exist among all children. Targeted interventions will benefit from mechanisms to identify children with increased risks such as sickle cell anemia or trait to improve pneumococcal vaccination coverage among these groups.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/145560/1/pbc27282.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/145560/2/pbc27282_am.pd

Receptor Tyrosine Kinases Fall into Distinct Classes Based on Their Inferred Signaling Networks

Although many anticancer drugs that target receptor tyrosine kinases (RTKs) provide clinical benefit, their long-term use is limited by resistance that is often attributed to increased abundance or activation of another RTK that compensates for the inhibited receptor. To uncover common and unique features in the signaling networks of RTKs, we measured time-dependent signaling in six isogenic cell lines, each expressing a different RTK as downstream proteins were systematically perturbed by RNA interference. Network models inferred from the data revealed a conserved set of signaling pathways and RTK-specific features that grouped the RTKs into three distinct classes: (i) an EGFR/FGFR1/c-Met class constituting epidermal growth factor receptor, fibroblast growth factor receptor 1, and the hepatocyte growth factor receptor c-Met; (ii) an IGF-1R/NTRK2 class constituting insulin-like growth factor 1 receptor and neurotrophic tyrosine receptor kinase 2; and (iii) a PDGFRβ class constituting platelet-derived growth factor receptor β. Analysis of cancer cell line data showed that many RTKs of the same class were coexpressed and that increased abundance of an RTK or its cognate ligand frequently correlated with resistance to a drug targeting another RTK of the same class. In contrast, abundance of an RTK or ligand of one class generally did not affect sensitivity to a drug targeting an RTK of a different class. Thus, classifying RTKs by their inferred networks and then therapeutically targeting multiple receptors within a class may delay or prevent the onset of resistance.W. M. Keck FoundationNational Institutes of Health (U.S.) (R21 CA126720)National Institutes of Health (U.S.) (P50 GM068762)National Institutes of Health (U.S.) (RC1 HG005354)National Institutes of Health (U.S.) (U54-CA112967)National Institutes of Health (U.S.) (R01-CA096504)Alfred and Isabel Bader (Fellowship)Jacques-Emile Dubois (fellowship

Archaeology of the Planned Location of the Toyota Motor Manufacturing Plant, San Antonio, Bexar County, Texas

From October 2002 to January 2004, the Center for Archaeological Research (CAR) of The University of Texas at San Antonio conducted archaeological investigations for the City of San Antonio in a 2,570.25-acre project area that is the future site of the San Antonio Toyota Motor Manufacturing Plant. The work was conducted under Texas Antiquities Permit No. 2982 with Dr. Steve A. Tomka, CAR Director, serving as Principal Investigator. The project included the reconnaissance of over 500 acres of the project area, the excavation of 376 shovel tests, 250 mechanical auger borings, and 42 backhoe and Gradall trenches. The backhoe and Gradall trenches were dug for geoarchaeological investigations and in one instance to search for a presumed historic cemetery. Reassessment for National Register of Historic Places and State Archeological Landmark status was conducted for 16 previously documented archaeological sites (41BX125, 41BX349, 41BX652, 41BX653, 41BX654, 41BX655, 41BX656, 41BX657, 41BX658, 41BX659, 41BX660, 41BX661, 41BX662, 41BX676, 41BX681, and 41BX832) and five newly identified sites (41BX1571–41BX1575). Of the 21 sites examined during this project, 12 are prehistoric, seven are historic and two have both prehistoric and historic components. The prehistoric sites are lithic and burned rock scatters, possibly the remnants of campsites. Diagnostic artifacts found in previous surveys indicate Archaic and Late Prehistoric time frames. The historic sites present are farmstead-ranch complexes including residential structures and outbuildings. Also encountered were tenant farmer residences and a small brick kiln. The historic components are primarily late-nineteenth and early-twentieth-century, although original surveys noted early-nineteenth-century artifacts. All artifacts collected are curated at the Center for Archaeological Research laboratory facility

Profiling of Haemophilus influenzae strain R2866 with carbohydrate-based covalent probes.

We demonstrate the application of four covalent probes based on anomerically pure d-galactosamine and d-glucosamine scaffolds for the profiling of Haemophilus influenzae strain R2866. The probes have been used successfully for the labelling of target proteins not only in cell lysates, but also in intact cells. Differences in the labelling patterns between lysates and intact cells indicate that the probes can penetrate into the periplasm, but not the cytoplasm of H. influenzae. Analysis of selected target proteins by LC-MS/MS suggests predominant labelling of nucleotide-binding proteins, including several known antibacterial drug targets. Our protocols will aid the identification of molecular determinants of bacterial pathogenicity in Haemophilus influenzae and other bacterial pathogens