Nicolas Huot

Programmable focal spot shaping of amplified femtosecond laser pulses

We describe the programmable spatial beam shaping of 100-kHz, 4-microJ amplified femtosecond pulses in a focal plane by wave-front modulation. Phase distributions are determined by a numerical iterative procedure. A nonpixelated optically addressed liquid-crystal light valve is used as a programmable wave-front tailoring device. Top-hat, doughnut, square, and triangle shapes of 20-microm size are obtained in a focal plane. Their suitability for femtosecond laser machining is demonstrated.

Meganuclease-mediated Inhibition of HSV1 Infection in Cultured Cells

Herpes simplex virus type 1 (HSV1) is a major health problem. As for most viral diseases, current antiviral treatments are based on the inhibition of viral replication once it has already started. As a consequence, they impair neither the viral cycle at its early stages nor the latent form of the virus, and thus cannot be considered as real preventive treatments. Latent HSV1 virus could be addressed by rare cutting endonucleases, such as meganucleases. With the aim of a proof of concept study, we generated several meganucleases recognizing HSV1 sequences, and assessed their antiviral activity in cultured cells. We demonstrate that expression of these proteins in African green monkey kidney fibroblast (COS-7) and BSR cells inhibits infection by HSV1, at low and moderate multiplicities of infection (MOIs), inducing a significant reduction of the viral load. Furthermore, the remaining viral genomes display a high rate of mutation (up to 16%) at the meganuclease cleavage site, consistent with a mechanism of action based on the cleavage of the viral genome. This specific mechanism of action qualifies meganucleases as an alternative class of antiviral agent, with the potential to address replicative as well as latent DNA viral forms.

Analysis of the effects of spherical aberration on ultrafast laser-induced refractive index variation in glass

We propose a comprehensive analysis of the effects that spherical aberration may have on the process of ultrafast laser photowriting in bulk transparent materials and discuss the consequences for the generated refractive index changes. Practical aspects for a longitudinal photowriting configuration are emphasized. Laser-induced index variation in BK7 optical glass and fused silica (a-SiO(2)) affected by spherical aberration are characterized experimentally using phase-contrast optical microscopy. Experimental data are matched by analytical equations describing light propagation through dielectric interfaces. Corrective solutions are proposed with a particular focus on the spatial resolution achievable and on the conditions to obtain homogeneously photo-induced waveguides in a longitudinal writing configuration.

Characterization of a photorefractive rhodium doped barium titanate at 1.06 μm

Abstract We characterize Rh:BaTiO 3 by cw two-beam coupling experiments at 1.06 μm. We demonstrate that all our measurements are well described by a single-carrier, single-site model as soon as the incident intensity is large enough. The effective trap density is found to be equal to N eff = 6 × 10 16 cm −3 and the maximum measured gain is 23 cm −1 . At 20 W cm −2 with a measured absorption of 0.15 cm −1 , the time constant is 75 s.

Natural killer cells migrate into and control simian immunodeficiency virus replication in lymph node follicles in African green monkeys

Natural killer (NK) cells play an essential role in antiviral immunity, but knowledge of their function in secondary lymphoid organs is incomplete. Lymph node follicles constitute a major viral reservoir during infections with HIV-1 and simian immunodeficiency virus of macaques (SIVmac). In contrast, during nonpathogenic infection with SIV from African green monkeys (SIVagm), follicles remain generally virus free. We show that NK cells in secondary lymphoid organs from chronically SIVagm-infected African green monkeys (AGMs) were frequently CXCR5+ and entered and persisted in lymph node follicles throughout the follow-up (240 d post-infection). These follicles were strongly positive for IL-15, which was primarily presented in its membrane-bound form by follicular dendritic cells. NK cell depletion through treatment with anti-IL-15 monoclonal antibody during chronic SIVagm infection resulted in high viral replication rates in follicles and the T cell zone and increased viral DNA in lymph nodes. Our data suggest that, in nonpathogenic SIV infection, NK cells migrate into follicles and play a major role in viral reservoir control in lymph nodes.

Two-wave mixing in photorefractive BaTiO 3 :Rh at 1.06 µm in the nanosecond regime

We present two-beam coupling experiments in the nanosecond regime at 1.06 mum , using photorefractive BaTiO(3):Rh. The maximum observed exponential gain coefficient is 14.2 cm(-1) . No intensity-dependent electron-hole competition and no strong saturation of the photoionized charge carriers are observed for intensities of less than 20MW cm(-2) . The energy required for recording the photorefractive grating is not significantly different in the nanosecond and the cw regimes.

Spatial mode control of a diode-pumped Nd:YAG laser by use of an intracavity holographic phase plate

We present a new method of realizing phase plates by phase-volume holography on a photopolymer film. We implement such a component in a diode-pumped Nd:YAG oscillator to control the output spatial beam profile. Flattop super-Gaussian and square-shaped beams are obtained.

Self-pumped phase conjugation in a ring cavity at 1.06 μm in cw and nanosecond regimes using photorefractive BaTiO3:Rh

Abstract We present self-pumped phase conjugation experiments in a ring cavity at 1.06 μm in a BaTiO 3 :Rh crystal both in the cw and in the nanosecond regime. The measured reflectivities are respectively 50% and 66%. At 4 W cm −2 of cw illumination, it only takes 90 s to reach the steady state. A preliminary experiment of correction of the focusing effect of a lens is reported.

Plasmacytoid Dendritic Cell Infection and Sensing Capacity during Pathogenic and Nonpathogenic Simian Immunodeficiency Virus Infection

ABSTRACT Human immunodeficiency virus (HIV) in humans and simian immunodeficiency virus (SIV) in macaques (MAC) lead to chronic inflammation and AIDS. Natural hosts, such as African green monkeys (AGM) and sooty mangabeys (SM), are protected against SIV-induced chronic inflammation and AIDS. Here, we report that AGM plasmacytoid dendritic cells (pDC) express extremely low levels of CD4, unlike MAC and human pDC. Despite this, AGM pDC efficiently sensed SIVagm, but not heterologous HIV/SIV isolates, indicating a virus-host adaptation. Moreover, both AGM and SM pDC were found to be, in contrast to MAC pDC, predominantly negative for CCR5. Despite such limited CD4 and CCR5 expression, lymphoid tissue pDC were infected to a degree similar to that seen with CD4+ T cells in both MAC and AGM. Altogether, our finding of efficient pDC infection by SIV in vivo identifies pDC as a potential viral reservoir in lymphoid tissues. We discovered low expression of CD4 on AGM pDC, which did not preclude efficient sensing of host-adapted viruses. Therefore, pDC infection and efficient sensing are not prerequisites for chronic inflammation. The high level of pDC infection by SIVagm suggests that if CCR5 paucity on immune cells is important for nonpathogenesis of natural hosts, it is possibly not due to its role as a coreceptor. IMPORTANCE The ability of certain key immune cell subsets to resist infection might contribute to the asymptomatic nature of simian immunodeficiency virus (SIV) infection in its natural hosts, such as African green monkeys (AGM) and sooty mangabeys (SM). This relative resistance to infection has been correlated with reduced expression of CD4 and/or CCR5. We show that plasmacytoid dendritic cells (pDC) of natural hosts display reduced CD4 and/or CCR5 expression, unlike macaque pDC. Surprisingly, this did not protect AGM pDC, as infection levels were similar to those found in MAC pDC. Furthermore, we show that AGM pDC did not consistently produce type I interferon (IFN-I) upon heterologous SIVmac/HIV type 1 (HIV-1) encounter, while they sensed autologous SIVagm isolates. Pseudotyping SIVmac/HIV-1 overcame this deficiency, suggesting that reduced uptake of heterologous viral strains underlays this lack of sensing. The distinct IFN-I responses depending on host species and HIV/SIV isolates reveal the host/virus species specificity of pDC sensing.

Structural Variability of the Herpes Simplex Virus 1 Genome In Vitro and In Vivo

ABSTRACT Herpes simplex virus 1 (HSV-1) is a human pathogen that leads to recurrent facial-oral lesions. Its 152-kb genome is organized in two covalently linked segments, each composed of a unique sequence flanked by inverted repeats. Replication of the HSV-1 genome produces concatemeric molecules in which homologous recombination events occur between the inverted repeats. This mechanism leads to four genome isomers (termed P, IS, IL, and ILS) that differ in the relative orientations of their unique fragments. Molecular combing analysis was performed on DNA extracted from viral particles and BSR, Vero, COS-7, and Neuro-2a cells infected with either strain SC16 or KOS of HSV-1, as well as from tissues of experimentally infected mice. Using fluorescence hybridization, isomers were repeatedly detected and distinguished and were accompanied by a large proportion of noncanonical forms (40%). In both cell and viral-particle extracts, the distributions of the four isomers were statistically equivalent, except for strain KOS grown in Vero and Neuro-2a cells, in which P and IS isomers were significantly overrepresented. In infected cell extracts, concatemeric molecules as long as 10 genome equivalents were detected, among which, strikingly, the isomer distributions were equivalent, suggesting that any such imbalance may occur during encapsidation. In vivo, for strain KOS-infected trigeminal ganglia, an unbalanced distribution distinct from the one in vitro was observed, along with a considerable proportion of noncanonical assortment.