Imaging interactions between the immune and cardiovascular systems in vivo by multiphoton microscopy

Several recent studies in immunology have used multiphoton laser-scanning microscopy to visualise the induction of an immune response in real time in vivo. These experiments are illuminating the cellular and molecular interactions involved in the induction, maintenance and regulation of immune responses. Similar approaches are being applied in cardiovascular research where there is an increasing body of evidence to support a significant role for the adaptive immune system in vascular disease. As such, we have begun to dissect the role of T lymphocytes in atherosclerosis in real time in vivo. Here, we provide step-by-step guides to the various stages involved in visualising the migration of T cells within a lymph node and their infiltration into inflamed tissues such as atherosclerotic arteries. These methods provide an insight into the mechanisms involved in the activation and function of immune cells in vivo

Donor species complement after liver xenotransplantation: The mechanism of protection from hyperacute rejection

Hamster hearts transplanted into stable rat recipients of hamster livers (OLT rats) were hyperacutely rejected after transfer with unaltered rat antihamster hyperimmune serum (HS). This was followed by immediate liver xenograft rejection in 4 of 5 rats. In contrast, simple heat inactivation of the rat HS resulted in prolonged survival of hamster hearts to 25 days without deterioration effect in the liver xenografts. This effect was species-specific because third-party mouse heart grafts in OLT rats were hyperacutely rejected in minutes if either active or heat inactivated antimouse HS was given. In cytotoxicity experiments, the complement in OLT serum produced weak lysis of hamster lymphocytes, while efficiently doing so with mouse cell targets. Because normal hamster serum caused no lysis at all of hamster target cells, the residual low-grade lysis of OLT serum was possibly being mediated by extrahepatic sources of rat C. In conclusion, the homology of C and target cells represents a mechanism of protection that the liver confers to other organs, and that is most easily seen in xenografts but may be allospecifically operational with allografts as well within the limits of MHC restriction. © 1994 by Williams and Wilkins

Resonant Processes in a Frozen Gas

We present a theory of resonant processes in a frozen gas of atoms interacting via dipole-dipole potentials that vary as $r^{-3}$, where $r$ is the interatomic separation. We supply an exact result for a single atom in a given state interacting resonantly with a random gas of atoms in a different state. The time development of the transition process is calculated both on- and off-resonance, and the linewidth with respect to detuning is obtained as a function of time $t$. We introduce a random spin Hamiltonian to model a dense system of resonators and show how it reduces to the previous model in the limit of a sparse system. We derive approximate equations for the average effective spin, and we use them to model the behavior seen in the experiments of Anderson et al. and Lowell et al. The approach to equilibrium is found to be proportional to $\exp (-\sqrt{\gamma_{eq}t}$), where the constant $\gamma _{eq}$ is explicitly related to the system's parameters.Comment: 30 pages, 6 figure

Stability of a buoyant Oldroyd-B flow saturating a vertical porous layer with open boundaries

The performance of several engineering applications are strictly connected to the rheology of the working fluids and the Oldroyd-B model is widely employed to describe a linear viscoelastic behaviour. In the present paper, a buoyant Oldroyd-B flow in a vertical porous layer with permeable and isothermal boundaries is investigated. Seepage flow is modelled through an extended version of Darcy’s law which accounts for the Oldroyd-B rheology. The basic stationary flow is parallel to the vertical axis and describes a single-cell pattern where the cell has an infinite height. A linear stability analysis of such a basic flow is carried out to determine the onset conditions for a multicellular pattern. This analysis is performed numerically by employing the shooting method. The neutral stability curves and the values of the critical Rayleigh number are evaluated for different retardation time and relaxation time characteristics of the fluid. The study highlights the extent to which the viscoelasticity has a destabilising effect on the buoyant flow. For the limiting case of a Newtonian fluid, the known results available in the literature are recovered, namely a critical value of the Darcy–Rayleigh number equal to 197.081 and a corresponding critical wavenumber of 1.05950

Humoral and cellular immunopathology of hepatic and cardiac hamster-into-rat xenograft rejection: Marked stimulation of IgM^++bright/IgD^+dull splenic B cells

Normal Lewis rat serum contains antibodies (IgM > IgG) that bind to hamster leukocytes and endothelial cells. Transplantation of either the heart or liver from hamster rat results in release of hamster hematolymphoid cells from the graft, which lodge in the recipient spleen (cell migration), where recipient T- and B-cell populations initiate DNA synthesis within one day. There is marked stimulation of splenic IgM++(bright)/IgD+(dull) B cells in the marginal zone and red pulp, which account for 48% of the total splenic blast cell population by 4 days after liver transplantation. CD4+ predominant T-cell proliferation in the splenic periarterial lymphatic sheath and paracortex of peripheral lymph nodes occurs almost simultaneously. The effector phase of rejection in cardiac recipients is dominated by complement-fixing IgM antibodies, which increase daily and result in graft destruction in 3 to 4 days, even in animals treated with FK506. In liver recipients, combined antibody and cellular rejection, associated with graft infiltration by OX8+ natural killer, and fewer W3/25+ (CD4) lymphocytes, are responsible for graft failure in untreated recipients at 6 to 7 days. FK506 inhibits the T-cell response in liver recipients and significantly prolongs graft survival, but does not prevent the rise or deposition of IgM antibodies in the graft. However, a single injection of cyclophosphamide 10 days before transplantation effectively depletes the splenic IgM++(bright)/IgD+(dull) cells and in combination with FK506, results in 100% survival of both cardiac and hepatic xenografts for more than 60 days. Although extrapolation of morphological findings to functional significance is fraught with potential problems, we propose the following mechanisms of xenograft rejection. The reaction initially appears to involve primitive host defense mechanisms, including an IgM-producing subpopulation of splenic B cells and natural killer cells. Based on the reaction and distribution of OX8+ and W3/25+ cells, antibody dependent cell cytotoxicity and delayed-type hypersensitivity responses seem worthy of further investigation as possible effector mechanisms. Effective control of xenograft rejection is likely to require a dual pharmaceutical approach, one to contain T-cell immunity and another to blunt the primitive B-cell response