Induction of specific tolerance by intrathymic injection of recipient muscle cells transfected with donor class I major histocompatibility complex.

Induction of tolerance to allogeneic MHC antigens has been a goal in the field of transplantation because it would reduce or eliminate the need for generalized immunosuppression. Although encouraging results have been obtained in experimental models by exposing recipient thymus to donor cells before transplantation, donor cells are not typically available at that time, and the donor antigens responsible for the effect are poorly defined. In the present study, thymic tolerance was demonstrated without using donor cells. Recipient thymus was injected before transplantation with autologous myoblasts and myotubes that were genetically modified to express allogeneic donor-type MHC class I antigen. Donor-specific unresponsiveness was induced to a completely MHC-disparate liver transplant and to a subsequent donor-type cardiac allograft, but not a third-party allograft. In vitro, recipient CTL demonstrated a 10-fold reduction in killing of donor cells, but not of third-party cells. Our results demonstrate: (1) that recipient muscle cells can be genetically engineered to induce donor-specific unresponsiveness when given intrathymically, and (2) transfected recipient cells expressing only donor MHC class I antigen can induce tolerance to a fully allogeneic donor

Immunity to MHC class I antigen after direct DNA transfer into skeletal muscle.

Plasmid cDNA encoding the alpha-chain of either membrane-bound (pcRT.45) or secreted (pcRQ.B3) RT1Aa MHC class I Ag were transferred to Lewis (RT1(1)) rat skeletal muscle by direct injection. Rats were challenged 7 days later with an ACI (RT1a) heterotropic heart transplant, and cardiac allograft survival, RT1Aa-specific antibody levels, and frequency of ACI-specific CTL were monitored. Graft rejection was accelerated by > or = 2 days in an Ag-specific and dose-dependent manner in pcRT.45-injected rats. The pcRQ.B3-injected rats also rejected grafts more rapidly; however, graft rejection was accelerated by only 1 day, and graft infiltrates were less pronounced than in pcRT.45-injected rats. Injection of pcRT.45 resulted in an increase in ACI-specific CTL precursor frequency 3 days post-transplant, whereas there was no significant change in rats pretreated with pcRQ.B3 injection. Compared with rats injected with a control plasmid encoding firefly luciferase, transfer of pcRT.45 resulted in an increase in RT1Aa-specific IgG and IgM antibody 3 days after heart transplantation. Transfer of pcRQ.B3 resulted in a similar mean increase in RT1Aa-specific IgG and IgM antibody after transplantation, but the variability from rat to rat was greater, with some animals exhibiting strong priming, and others showing little or no priming by gene injection. Our results suggest that skeletal muscle can express either membrane-bound or secreted MHC class I Ag after gene transfer, but that the membrane-bound form is more immunogenic than the secreted form in the high responder Lewis rat. Direct DNA transfer to skeletal muscle provides a rapid and specific approach to studying immunity to allogeneic MHC Ag

Use of donor serum to prevent passive transfer of hyperacute rejection

Organ transplantation in presensitized recipients continues to be contraindicated for heart and kidney recipients due to the risk of hyperacute rejection, which has no known treatment at this time. We tested whether donor serum, which contains soluble MHC class I antigen, is able to neutralize the effect of anti-donor antibody in the recipient and prevent hyperacute or accelerated rejection. A rat model of passive immunization was used to test the role of anti-donor antibody in hyperacute rejection. Seven of 10 recipients of hyperimmune serum (HyS), derived from Lewis rats (RT1l) following 3 ACI (RT1a) skin grafts, developed hyperacute or accelerated rejection. Intravenous injection of ACI serum prior to the HyS administration prevented hyperacute rejection in all recipients tested. When third-party (Wistar-Furth, RT1u) serum was given to Lewis rats injected with HyS, hyperacute rejection was not abrogated. When examining the mechanism of this effect, a simple antibody blocking phenomenon was found to be unlikely since flow cytometry analysis showed that ACI serum needed to be present at > or = 256-fold excess compared to HyS to block anti-ACI antibody binding to RT1.Aa+cells by 50%. We tested whether the RT1.Aa class I antigen in ACI serum had other biologic properties that resulted in the prolonged graft survival. However, removal of RT1.Aa antigen from ACI serum prior to use in the passive transfer model did not abrogate the graft prolongation observed previously. These data suggest that components of donor serum other than MHC class I antigen may be useful for preventing the antibody-mediated component of hyperacute rejection

Discovery of pulsations in the X-ray transient 4U 1901+03

We describe observations of the 2003 outburst of the hard-spectrum X-ray transient 4U 1901+03 with the Rossi X-ray Timing Explorer. The outburst was first detected in 2003 February by the All-Sky Monitor, and reached a peak 2.5-25 keV flux of 8x10^-9 ergs/cm^2/s (around 240 mCrab). The only other known outburst occurred 32.2 yr earlier, likely the longest presently known recurrence time for any X-ray transient. Proportional Counter Array (PCA) observations over the 5-month duration of the 2003 outburst revealed a 2.763 s pulsar in a 22.58 d orbit. The detection of pulsations down to a flux of 3x10^-11 ergs/cm^2/s (2.5-25 keV), along with the inferred long-term accretion rate of 8.1x10^-11 M_sun/yr (assuming a distance of 10 kpc) suggests that the surface magnetic field strength is below ~5x10^11 G. The corresponding cyclotron energy is thus below 4 keV, consistent with the non-detection of resonance features at high energies. Although we could not unambiguously identify the optical counterpart, the lack of a bright IR candidate within the 1' RXTE error circle rules out a supergiant mass donor. The neutron star in 4U 1901+03 probably accretes from the wind of a main-sequence O-B star, like most other high-mass binary X-ray pulsars. The almost circular orbit e=0.036 confirms the system's membership in a growing class of wide, low-eccentricity systems in which the neutron stars may have received much smaller kicks as a result of their natal supernova explosions.Comment: 7 pages, 6 figures, accepted by ApJ. Very minor addition in response to referee's comment; updated author affiliatio

HCMV-encoded NK modulators: Lessons from in vitro and in vivo genetic variation

Human cytomegalovirus (HCMV) is under constant selective pressure from the immune system in vivo. Study of HCMV genes that have been lost in the absence of, or genetically altered by, such selection can focus research toward findings of in vivo significance. We have been particularly interested in the most pronounced change in the highly passaged laboratory strains AD169 and Towne—the deletion of 13–15 kb of sequence (designated the UL/b′ region) that encodes up to 22 canonical genes, UL133-UL150. At least 5 genes have been identified in UL/b′ that inhibit NK cell function. UL135 suppresses formation of the immunological synapse (IS) by remodeling the actin cytoskeleton, thereby illustrating target cell cooperation in IS formation. UL141 inhibits expression of two activating ligands (CD155, CD112) for the activating receptor CD226 (DNAM-1), and two receptors (TRAIL-R1, R2) for the apoptosis-inducing TRAIL. UL142, ectopically expressed in isolation, and UL148A, target specific MICA allotypes that are ligands for NKG2D. UL148 impairs expression of CD58 (LFA-3), the co-stimulatory cell adhesion molecule for CD2 found on T and NK cells. Outside UL/b′, studies on natural variants have shown UL18 mutants change affinity for their inhibitory ligand LIR-1, while mutations in UL40's HLA-E binding peptide differentially drive NKG2C+ NK expansions. Research into HCMV genomic stability and its effect on NK function has provided important insights into virus:host interactions, but future studies will require consideration of genetic variability and the effect of genes expressed in the context of infection to fully understand their in vivo impact

Coherent particle production in collisions of relativistic nuclei

Here we give the results of our study of features of dense groups, or spikes, of particles produced in Mg-Mg and C-Cu collisions at, respectively, 4.3 and 4.5 GeV/c/nucleon aimed to search for a coherent, Cerenkov-like, mechanism of hadroproduction. We investigate the distributions of spike centers and, for Mg-Mg interactions, the energy spectra of negatively charged particles in spikes. The spike-center distributions are obtained to exhibit the structure expected from coherent gluon-jet emission dynamics. This structure is similar in both cases considered, namely for all charged and negatively charged particles, and is also similar to that observed recently for all-charged-particle spikes in hadronic interactions. The energy distribution within spikes is found to have a significant peak over the inclusive background, while the inclusive spectrum shows exponential decrease with two characteristic values of average kinetic energy. The value of the peak energy and its width are in a good agreement with those expected for pions produced in a nuclear medium in the framework of the Cerenkov quantum approach. The peak energy obtained is consistent with the value of the cross-section maximum observed in coincidence nucleon-nucleus interaction experiments.Comment: 8 pages, 5 figures. Invited talk presented by E.S. at the 9th International Workshop on Multiparticle Production: New Frontiers in Soft Physics and Correlations on the Threshold of the Third Millenium, Turin, Italy, June 12 - 17, 200

Photodynamic priming with triple-receptor targeted nanoconjugates that trigger T cell-mediated immune responses in a 3D in vitro heterocellular model of pancreatic cancer

Photodynamic priming (PDP), a collateral effect of photodynamic therapy, can transiently alter the tumor microenvironment (TME) beyond the cytotoxic zone. Studies have demonstrated that PDP increases tumor permeability and modulates immune-stimulatory effects by inducing immunogenic cell death, via the release of damage-associated molecular patterns and tumor-associated antigens. Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest of cancers with a stubborn immunosuppressive TME and a dense stroma, representing a challenge for current molecular targeted therapies often involving macromolecules. We, therefore, tested the hypothesis that PDP\u27s TME modulation will enable targeted therapy and result in immune stimulation. Using triple-receptor-targeted photoimmuno-nanoconjugate (TR-PINs)-mediated PDP, targeting epidermal growth factor receptor, transferrin receptor, and human epidermal growth factor receptor 2 we show light dose-dependent TR-PINs mediated cytotoxicity in human PDAC cells (MIA PaCa-2), co-cultured with human pancreatic cancer-associated fibroblasts (PCAFs) in spheroids. Furthermore, TR-PINs induced the expression of heat shock proteins (Hsp60, Hsp70), Calreticulin, and high mobility group box 1 in a light dose and time-dependent manner. TR-PINs-mediated T cell activation was observed in co-cultures of immune cells with the MIA PaCa-2-PCAF spheroids. Both CD4+ T and CD8+ T cells showed light dose and time-dependant antitumor reactivity by upregulating degranulation marker CD107a and interferon-gamma post-PDP. Substantial tumor cell death in immune cell-spheroid co-cultures by day 3 shows the augmentation by antitumor T cell activation and their ability to recognize tumors for a light dose-dependent kill. These data confirm enhanced destruction of heterogeneous pancreatic spheroids mediated by PDP-induced phototoxicity, TME modulation and increased immunogenicity with targeted nanoconstructs

AN INNOVATIVE LEE, MASSACHUSETTS USA DISSOLVED AIR FLOTATION POTABLE WATER FILTRATION PLANT

Wang, LK, Wang, MHS and Fahey, E (2020). An innovative Lee, Massachusetts USA dissolved air flotation potable water filtration plant. In: "Evolutionary Progress in Science, Technology, Engineering, Arts, and Mathematics (STEAM)", Wang, Lawrence K. and Tsao, Hung-ping (editors). Volume 2, Number 1, January 2020; 60 pages. Lenox Institute Press, Newtonville, NY, 12128-0405, USA. No. STEAM-VOL2-NUM1-JAN2020; ISBN 978-0-9890870-3-2---------------The authors present the overall structural design, design criteria, and performance data of the two Krofta Sandfloat flotation-filtration clarifiers (DAFF; SAF-BP24) installed at the 2.0 MGD (7570 m3/day) Lee Plant in treatment of surface water for potable purposes. Lenox Institute of Water Technology (LIWT) invented and patented the innovative DAFF system, while Krofta Engineering Corporation (KEC) manufactured and installed the Lee Plant. The author also discuss (a) current corrosion control program in order to comply with the US Federal Copper and Lead Rule, and (b) the current DAF-filtration plant’s performance for removal of perchlorate, barium, sodium, disinfection by-products (DBP), total trihalomethane (THM), total haloacetic acid (HAA), microbial contaminants, turbidity, iron and manganese. The 19 years old innovative Lee DAF-filtration plant met all US Environmental Protection Agency, and the Commonwealth of Massachusetts primary and secondary drinking water standards in accordance with the 2018 Water Quality Report. The Town of Lee successfully uses zinc orthophosphate and pH adjustment to stabilize the water throughout of Lee distribution system for lead, copper and pipe corrosion control. This article has been written in memory of Dr. Milos Krofta, Dr. Donald B. Aulenbach and Dr. William A. Selke