Ontogenic Development of Th1 and Th2 Cytokine Capabilities in Random Bred Mice

Neonatal mouse Th1 capabilities mature by postnatal day 5. Neonatal T cells have been reported to exhibit a bias towards Th2 cytokine production when co-cultured with adult antigen presenting cells (APC). We studied mouse T cells co-cultured with contemporary APC to evaluate neonatal cytokine production capabilities. In response to allogeneic stimulation, T cells co-cultured with contemporary APC from day 5 pups produced 37-fold greater IFNγ and 1.4-fold greater IL-2 levels than day 20 weanling mice. After CD3 ligation, cells from day 5 pups produced 4- (IL-2) and 10-fold (IFNγ) greater levels than adults (day 45), and concentrations were 27- (IL-2) and 18-fold (IFNγ) higher than with allogeneic stimulation alone. On average, the percent difference in concentrations was 418 (IL-4), 286 (IL-2) and 1140% (IFNγ) higher in unseparated spleen cells than in isolated splenic CD4 cells and APC. These results demonstrate that, in response to allogeneic stimulation with or without CD3 ligation, lymphocytes of neonatal mice (day 5) have the capacity to produce equivalent or greater TcR-dependent Th1 cytokine (IL-2 and IFNγ) levels than adult mice. Findings also support the idea that the reported Th2 bias of neonatal T cells may be the result of in vitro manipulation and choice of mouse strain, not of an inherent bias

Maternal Modulation of Neonatal Immune System Development

Changes in programming of neonatal immune development were effected through maternal immune modulation (Leishmania major inoculation). In progeny of these dams, immune profiles in both blood and spleen were changed throughout the neonatal period and were pronounced after weaning. White blood cell (WBC) and lymphocyte counts in blood of 45-day-old progeny were two-fold less than control animals. In blood, proportions of B cells were greater, while T helpers, Tc/s and NK cells were less than in controls. In contrast, proportions of splenic B and NK cells were greater than controls. But, proportions of all T and Tc/s cells on d20 and 45 were lower than controls. In blood, absolute numbers of all T, Th naïve and Th memory cells were lower than in controls. In contrast, in the spleen, numbers of NK, T and Th naïve and memory cells were up to 200% greater than in control pups. Cytokine responses of splenic lymphocytes stimulated through CD3 ligation revealed no difference in IL-4 production. In contrast, IL-2 and IFNγ were lower on d45 and 5, respectively, in the experimental compared to control mice. These data support the hypothesis that maternal immune events during gestation can modulate the pattern of immune development in offspring

Maturation of Lymphocyte Immunophenotypes and Memory T Helper Cell Differentiation During Development in Mice

The goal of this study was to systematically investigate the ontogeny of lymphoid populations throughout postnatal development. In CD-1 mice, peak lymphocyte numbers occurred in blood on postnatal day 10 (dl0) including those for natural killers (NK1.1), B cells (CD19), T helper (CD3CD4), naïve T helper (CD4CD62L(pos)CD44(low)), memory T helper (CD4CD62L(neg)CD44(high)), and T cytotoxic (CD3CD8) cells. As percent of total lymphocytes, peaks were achieved by d10 for all T helper subtypes but not B cells which declined to a nadir. In spleen, lymphocyte numbers increased exponentially after d10. Proportionately, NK and T cells peaked on d10, declined by d20, and increased 2–3-fold by d45. Naive T cells constituted the majority of lymphocytes during development while memory cells gained to 2.2% (blood) and 12 % (spleen) by d20. C57BL/6 mice had similar profiles except that the B cell nadir and T cell subset peaks were at d5. Peripheralization of critical numbers of lymphocytes by d10, and importantly, development of a repertoire of memory cells by d20, may define immune response capabilities that close the period of immaturity for the neonate

Engineering of gibberellin levels in citrus by sense and antisense

http://jxb.oxfordjournals.org/Carrizo citrange (Citrus sinensis x Poncirus trifoliata) is a citrus hybrid 4 widely used as a rootstock, whose genetic manipulation to improve 5 different growth characteristics is of high agronomic interest. In this work 6 we have produced transgenic Carrizo citrange plants overexpressing 7 sense and antisense CcGA20ox1 (a key enzyme of GA biosynthesis) 8 under control of the 35S promoter to modify plant architecture. As 9 expected, taller (sense) and shorter (antisense) phenotype correlated with 10 higher and lower levels, respectively, of active GA1 in growing shoots. In 11 contrast, other phenotypic characteristics seemed to be specific of citrus, 12 or different to those described for similar transgenics in other species. For 13 instance thorns, typical organs of citrus at juvenile stages, were much 14 longer in sense and shorter in antisense plants, and xylem tissue was 15 reduced in leaf and internode of sense plants. Antisense plants presented 16 a bushy phenotype, suggesting a possible effect of GAs on auxin 17 biosynthesis and/or transport. The main foliole of sense plants was longer, 18 although total leaf area was reduced. Leaf thickness was smaller in sense 19 and bigger in antisense plants due to changes in the spongy parenchyma. 20 Internode cell length was not altered in transgenic plants, indicating that in 21 citrus GAs regulate cell division rather than cell elongation. Interestingly, 22 the described phenotypes were not apparent when transgenic plants were 23 grafted on non-transgenic rootstock. This suggests that roots contribute to 24 the GA economy of aerial parts in citrus and opens the possibility of using 25 the antisense plants as dwarfing rootstocks.We thank J.A. Pina for technical assistance, and Dr. E. Carbonell and J. 34 Pérez for statistical analyses. This research was supported in part by grants CICYT AGL2003-01644, 1 INIA RTA04-13 and BIO2003-00151. C. 2 Fagoaga was recipient of an INIA-CCAA postdoctoral contract. I. Lliso was 3 recipient of an IVIA predoctoral fellowship. D.J. Iglesias and F.R. Tadeo 4 were recipients of INIA-CCAA and “Ramón y Cajal” MEC postdoctoral 5 contracts, respectively. 6 7Peer reviewe

Transgenic citrus plants expressing the citrus tristeza virus p23 protein exhibit viral-like symptoms

The 23 kDa protein (p23) coded by the 3'-terminal gene of Citrus tristeza virus (CTV), a member of the genus Closterovirus with the largest genome among plant RNA viruses, is an RNA-binding protein that contains a motif rich in cysteine and histidine residues in the core of a putative zinc-finger domain. On this basis, a regulatory role for CTV replication or gene expression has been suggested for p23. To explore whether over-expression of this protein in transgenic plants could affect the normal CTV infection process, transgenic Mexican lime plants were generated carrying the p23 transgene, or a truncated version thereof, under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Constitutive expression of p23 induced phenotypic aberrations that resembled symptoms incited by CTV in non-transgenic lime plants, whereas transgenic plants expressing the p23 truncated version were normal. The onset of CTV-like symptoms in p23 transgenic plants was associated with the expression of p23, and its accumulation level paralleled the intensity of the symptoms. This demonstrates that p23 is involved in symptom development and that it most likely plays a key role in CTV pathogenesis. This is the first case in which a protein encoded by a woody plant-infecting RNA virus has been identified as being directly involved in pathogenesis in its natural host. This finding also delimits a small region of the large CTV genome for the future mapping of specific pathogenic determinants

Latent Tuberculosis Infection in a Migrant Agricultural Community in Baja California, Mexico

The objectives were to estimate the prevalence and identify correlates of latent tuberculosis infection (LTBI) among residents of a migrant agricultural community in San Quintín, Baja-California, Mexico. Residents completed a questionnaire and had their blood tested for LTBI using the QuantiFERON®-TB Gold In-Tube (QFT) assay. Among 133 participants, 39.8% (95% CI 31.5–48.7%) tested QFT-positive. Having crossed the U.S.-Mexican border since living in San Quintin (P = 0.03), consuming unpasteurized milk (P = 0.02) and receiving health care at IMSS-Oportunidades in the last 6 months (P = 0.03) were independently associated with QFT-positivity. High LTBI prevalence in this community emphasizes the need for TB education and LTBI treatment for its residents. Association with travel to the U.S. suggests the potential for TB transmission across borders. Higher QFT-positivity among those consuming unpasteurized milk could indicate M. bovis infection, previously reported among Mexican migrants living in U.S. border cities

A multi-proxy approach to the palaecological reconstruction of the Orce Basin Archaeological Zone (Granada, Spain)

Comunicación oral presentada en XXI INQUA Congress. July 14th – 20th 2023, Rome (Italy)The Orce Basin Archaeological Zone (OZAB, Granada, Spain) extends over a surface area of some > 8.5 km2 and constitutes one of the richest Pleistocene vertebrate fossil records in western Europe including one of the oldest hominin presence in this part of Eurasia. Exceptionally rich collections of stone tools have been excavated from both of the Orce Oldowan sites: Barranco León (BL) (1.4 Ma) and Fuente Nueva 3 (FN3) (1.2 Ma), while BL has yielded a hominin deciduous lower molar. We present a multi-proxy approach to determine the palaeoeocological context of these first hominin settlements in Western Europe. By combining results from macrovertebrates, microvertebrates, as well as from pollen and stable isotopes from macrovertebrate tooth enamel, we determine whether the palaeo landscapes were dominated by savanna or open woodland. The results reveal the regional specificities of the OAZB, and also allow us to infer local features within the Orce sites. Overall, our data reveal the dominance of a typically Mediterranean climate and landscape since 1.8 Ma ago. The climatic conditions were generally more humid than at present, with warmer temperatures during the coldest months, indicating a higher net primary productivity (NPP). We find that precipitation and NPP appear to have been limiting factors for hominin presence in the OAZB. Thus, at the older palaeontological site of Venta Micena (1.6 Ma), climatic conditions appear to have been less compatible with hominin presence than during the BL and FN3 sequences, when early hominins inhabiting the OAZB were able to cope with changing climatic and environmental settings. Lastly, the comparison of the isotopic results of the Orce sites with those of the contemporaneous Shungura Formation (Ethiopia) reveals that the habitat in the westernmost part of Eurasia was distinctly unlike a typical African savanna

Genes involved in ethylene and gibberellins metabolism are required for endosperm-limited germiantion of Sisymbrium officinales L. Seeds

The rupture of the seed coat and that of the endosperm were found to be two sequential events in the germination of Sisymbrium officinale L. seeds, and radicle protrusion did not occur exactly in the micropylar area but in the neighboring zone. The germination patterns were similar both in the presence of gibberellins (GA4+7) and in presence of ethrel. The analysis of genes involved in GAs synthesis and breakdown demonstrated that (1) SoGA2ox6 expression peaked just prior to radicle protrusion (20–22 h), while SoGA3ox2 and SoGA20ox2 expression was high at early imbibition (6 h) diminishing sharply thereafter; (2) the accumulation of SoGA20ox2 transcript was strongly inhibited by paclobutrazol (PB) as well as by inhibitors of ET synthesis and signaling (IESS) early after imbibition (6 h), while SoGA3ox2 and SoGA2ox6 expression was slowly depressed as germination progressed; (3) ethrel and GA4+7 positively or negatively affected expression of SoGA3ox2, SoGA20ox2, and SoGA2ox6, depending on the germination period studied. Regarding genes involved in ET synthesis, our results showed that SoACS7 was expressed, just prior to radicle emergence while SoACO2 expression slowly increased as germination progressed. Both genes were strongly inhibited by PB but were almost unaffected by externally added ethrel or GA4+7. These results suggest that GAs are more important than ET during the early stages of imbibition, while ET is more important at the late phases of germination of S. officinale L. seed

Effect of Maternal HIV-1 Status and Antiretroviral Drugs on Haematological Profiles of South African Infants in Early Life

Maternal HIV-1 status and antiretroviral drug exposure may influence the haematological profiles of infants. We recruited infants from 118 uninfected control women and from 483 HIV-1 infected women who received no antiretroviral drugs (n=28), or received single-dose Nevirapine (sdNVP) (n=424) or triple-drug combination therapy (n=31) to reduce HIV-1 transmission. Blood was drawn from infants within 24 hours of delivery or 6-12 weeks post-delivery and full blood counts performed using a fully automated AcT-5-diff haematology analyser and reference controls. Exposed uninfected (EU; no NVP) differed from control infants only in having lower basophil counts and percentages. In all infant groups, leukocyte profiles showed characteristic quantitative changes with age in the first 6 weeks of life. HIV-1 infected infants displayed by 6 weeks elevations in white blood cells, lymphocyte, monocyte and basophil counts, and monocyte and basophil percentages, when compared to EU infants. At birth EU NVP-treated infants exhibited elevated monocyte percentages and counts and basophil counts that did not persist at 6 weeks. Interestingly, EU newborns of mothers with high CD4 counts (> 500 cells/μl) that had taken sdNVP had significantly elevated white blood cell, monocyte and basophil counts when compared to newborn infants of mothers with similar CD4 counts that had not taken sdNVP; this was not evident in infants of mothers with CD4 counts <200 cells/μl. These previously undescribed features may affect immune response capability in early life and clinical consequences of such changes need to be further investigated