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A simple method for unbiased quantitation of adoptively transferred cells in solid tissues.

In a mouse model, we demonstrate how to obtain a direct, unbiased estimate of the total number of adoptively transferred cells in a variety of organs at different time points. The estimate is obtained by a straightforward method based on the optical fractionator principle. Specifically, non-stimulated C57BL/6J mouse splenocytes were labelled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and adoptively transferred to normal C57BL/6J mice by intravenous injection. The total number of CFSE-positive cells was subsequently determined in lung, spleen, liver, kidney, and inguinal lymph node at six different time points following adoptive transfer (from 60 s to 1 week), providing a quantitative estimate of the organ distribution of the transferred cells over time. These estimates were obtained by microscopy of uniform samples of thick sections from the respective organs. Importantly, the samples were chosen and prepared in accordance with the optical fractionator principle. We demonstrate that the method is simple, precise, and well suited for quantitative immunological studies.

Adoptive Transfer↗

Adoptive transfer of NK 1.1+ lymphocytes in immune-mediated colitis: a pro-inflammatory or a tolerizing subgroup of cells?

UNLABELLED: T lymphocytes expressing NK1.1 marker (NKT) have been suggested to play crucial roles in immune modulation. AIM: To determine the role of NK1.1+ cells in induction and maintenance of pro-inflammatory and/or tolerizing responses. METHODS: Colitis was induced in C57/B6 donor mice by intracolonic instillation of trinitrobenzenesulfonic acid (TNBS). Donor mice received five oral doses of colonic proteins extracted from TNBS-colitis colonic wall. Depletion of NK1.1+ lymphocytes was performed before lymphocyte harvesting. Splenocytes were harvested and separated into T-cell subpopulations, and transplanted into recipient mice before intracolonic instillation of TNBS. Standard clinical, macroscopic, and microscopic scores, and intracellular staining, flow cytometry, and cytotoxicity assays were performed. RESULTS: The adoptive transfer of CD4+ and NK1.1+ cells harvested from tolerized mice markedly ameliorated the colitis in recipient mice. In contrast, the adoptive transfer of CD8+ and double negative lymphocytes failed to transfer the tolerance. Recipients of splenocytes from tolerized mice exhibited an increase in CD4+ IL4+/CD4+ IFNgamma+ ratio. In contrast, recipients of splenocytes from NK1.1-depleted-tolerized mice exhibited severe colitis with a significant decrease of the CD4+ IL4+/CD4+ IFNgamma+ ratio. However adoptive transfer of splenocytes from non-tolerized NKT-depleted mice led to an alleviation of colitis with a relative increase of the CD4+ IL4+/CD4+ IFNgamma+ ratio. CONCLUSIONS: NK1.1+ lymphocytes play a critical role in immune regulation. They may be accountable for an alteration of the inflammatory response and the CD4+ IL4+/CD4+ IFNgamma ratio immune-mediated colitis and in peripheral tolerance induction.

Adoptive Transfer↗

Complementary and alternative therapies: considerations for families after international adoption.

Children who are internationally adopted are at increased risk of developmental and behavioral concerns, including attention disorders, learning disorders, and autistic spectrum disorders. In attempting to promote their child's optimal development and well-being, parents of internationally adopted children are faced with the additional stress of having many unanswered and unanswerable questions about their child's early origins. As a result, internationally adopted children and their parents need the support and counsel of their pediatrician as they grow and develop into adulthood. A combination of traditional, complementary, and alternative therapies is the rule rather than the exception for most children with developmental challenges.

Adoption↗

Education and internationally adopted children: working collaboratively with schools.

Families who adopt internationally often need assistance in determining which factors they need to consider when making educational decisions for their child, and they frequently seek guidance from their physician. Understanding how the education system functions, how it differs from the medical system, and how children who have been adopted internationally can succeed in school is important for health care providers, because this information helps parents in making proactive decisions for their children. Internationally adopted children have the best chance of maximizing their learning potential when medical and educational professionals work together to assist families as they plan for their child's education.

Adoption↗

Synergistic effect of adoptive T-cell therapy and intratumoral interferon gamma-inducible protein-10 transgene expression in treatment of established tumors.

The lack of efficient T-cell infiltration of tumors is a major obstacle to successful adoptive T-cell therapy. We have previously shown that transplanted SP2/0 myeloma tumors engineered to express lymphotactin invariably induced tumor regress mediated by SP2/0 tumor-specific T cells. Herein, we further systemically characterize these activated T cells and investigate their therapeutic efficacy, either alone or with the chemokine interferon gamma (IFN-gamma)-inducible protein-10 (IP-10) gene therapy. Following stimulation with SP2/0 cells, these activated T cells were CD25(+)FasL(+) L-selectin(low), expressed CXCR3 receptor and were chemoattracted by IP-10 in vitro. They comprised 64% CD4(+) Th1 and 36% CD8(+) Tc1 cells, both of which expressed IFN-gamma, perforin, and TNF-alpha, but not IL-4. The activated T cells were strongly cytotoxic for SP2/0 tumor cells (79% specific killing; E:T ratio, 50), mainly via perforin-mediated pathway. Cell tracking using labeled T cells confirmed that these T cells infiltrated better into the IP-10-expressing tumors than non-IP-10-expressing ones. In vivo, combined intratumoral IP-10 gene transfer and adoptive T-cell immunotherapy for well-established SP2/0 tumors eradicated the tumors in 7 of the 8 mice. Control or IP-10 adenoviral treatments by themselves neither alter the lethal outcome for tumor-bearing mice nor did T-cell therapy by itself, although the latter two treatments did slow its time-frame. Taken together, our data provide solid evidence of a potent synergy between adoptive T-cell therapy and IP-10 gene transfer into tumor tissues, which culminated in the eradication of well-established tumor masses.

Adenoviridae↗

T-cell adoptive immunotherapy of metastatic renal cell carcinoma.

OBJECTIVES: To determine the feasibility and toxicity of the adoptive transfer of ex vivo-activated T lymphocytes that have been sensitized to autologous tumor vaccine in vivo. METHODS: Twenty patients with extensive metastatic renal cell carcinoma received systemic adoptive immunotherapy with autologous T cells in the absence of conjunctional interleukin-2 (IL-2) administration. Patients were vaccinated intradermally with irradiated autologous tumor cells and granulocyte-macrophage colony-stimulating factor as an adjuvant to stimulate an immune response. Inguinal lymph nodes draining the vaccine site were surgically removed, and the cells were stimulated with staphylococcal enterotoxin A followed by expansion in 60 IU/mL IL-2, and in some cases additionally stimulated with anti-CD3 monoclonal antibody and IL-2, to obtain a large number of cells. RESULTS: The staphylococcal enterotoxin A/IL-2 activation induced vigorous proliferation of T cells (median expansion 26-fold) that were a mixture of CD4 and CD8 T lymphocytes. Activated cells were infused intravenously at doses ranging from 2x10(9) to 9.5x10(10). There was minimal toxicity consisting of grade 1 or 2 fever and nausea, and the entire treatment was delivered as outpatient therapy. One patient had a partial response, one had a mixed response, and 8 had stable disease lasting at least 5 months. CONCLUSIONS: Adoptive transfer of ex vivo-activated, tumor vaccine-primed lymph node cells is feasible and is associated with minimal toxicity when used alone. These results warrant further study in a Phase II trial.

Adult↗

Suppression of human hepatoma in mice through adoptive transfer of immunity to the hepatitis B surface antigen.

BACKGROUND/AIMS: Adoptive transfer of immunity against hepatitis B surface antigen (HBsAg) has previously been shown to occur in mice and humans through transplantation of bone marrow cells from donors immunized against HBsAg (anti-HBs) to non-immune recipients. In the present study we evaluated the effect of adoptive transfer of immunity to HBsAg on the growth of HbsAg-secreting hepatocellular carcinoma (HCC) xenografts in athymic mice. METHODS: Immunocompetent mice were immunized with recombinant HBsAg. Bone marrow cells from anti-HBs+ mice were injected intravenously to irradiated athymic Balb/c mice which had been previously transplanted subcutaneously with Hep3B human hepatoma cells. Treatment groups included mice receiving bone marrow transplantation from HBV-immunized (anti-HBs positive) and non-immunized (anti-HBs negative) donors. RESULTS: At 9 weeks post bone marrow transplantation, tumor volume and serum alpha-fetoprotein levels in athymic mice receiving HBV-immune bone marrow cells were 11.5 mm3 and 363 ng/ml, respectively, as compared to 1579 mm3 and 19,000 ng/ml, in recipients of non-immune bone marrow transplantation (p<0.005). T-cell depletion of antiHBs+ immune bone marrow prior to transplantation decreased the anti-tumor effect but did not abolish it. A mild nonspecific, bone marrow-derived, graft versus tumor effect was observed in mice transplanted with human hepatoma cells that do not express HBsAg. CONCLUSIONS: Adoptive transfer of immunity to HBV facilitates suppression of experimental human HCC expressing HBsAg. This effect is the result of a combination of specific anti-viral surface antigen effect and a nonspecific graft versus tumor effect.

Adoptive Transfer↗

Unrepresentative behavior of T cell receptor-transgenic CD4+ T cells upon adoptive transfer: lack of need for priming and an extended booster dose-response.

The response of CD4+ T cells taken from DO11.10 alpha beta TCR-transgenic mice to their specific antigen, ovalbumin, was examined in an adoptive transfer system. Read out was the % frequency of KJI-26.1+ (clonotype positive) cells within the Thy-1.2+ (T cell) population in lymph nodes. Control experiments indicated that these cells were uniformly CD4+. Immunizing the transgenic mice had no detectable effect on this frequency. Furthermore, the frequency in recipients of adoptively transferred lymph node cells was not affected by priming of the donors with ovalbumin by various procedures. Transfers were into syngeneic SCID recipients, except in one experiment, where irradiated recipients were shown to behave in the same way. Examining the effect of varying the amount of booster antigen, the response increased slowly with dose, up to a plateau in the range of 10-100 mg ovalbumin. The lack of need for priming is unusual, in comparison with an adoptive transfer of non-transgenic cells, as is the extended dose response range with such a high optimum dose. This enhanced responsiveness is interpreted in terms of a lack of down-immunoregulation in these transgenic mice.

Adoptive Transfer↗

[Hepatitis A acquired from an asymptomatic adopted child].

We report the case of acute hepatitis in a 36-year-old woman that was acquired from an adopted African child with asymptomatic active infection. At present, most experts do not screen for hepatitis A. However, adoptive parents should be vaccinated against hepatitis A because of the risk of unrecognized active infection in adopted children from countries in which infection is endemic.

Acute Disease↗

Health care of the internationally adopted child. Part 2: Chronic care long-term medical issues.

Internationally adopted children remain at risk throughout their lives for medical sequelae related to their birth and residence in a foreign country. In the early period after placement the most important problems are the management of chronic infectious diseases and malnutrition. Over time, however, many children will have issues related to growth, age determination, timing of puberty, dental care, development, and language acquisition. Chronic disease and ethnic health considerations pose some special problems for the adopted child without a personal or family history. Part 1 of this series addressed the initial evaluation for the newly arrived internationally adopted child, whereas part 2 discusses the long-term management problems. J Pediatr Health Care.

Adolescent↗

CD4+CD25+ regulatory T cells generated in response to insulin B:9-23 peptide prevent adoptive transfer of diabetes by diabetogenic T cells.

NOD mice have a relative deficiency of CD4+CD25+ regulatory T cells that could result in an inability to maintain peripheral tolerance. The aim of this study was to induce the generation of CD4+CD25+ regulatory T cells in response to autoantigens to prevent type 1 diabetes (T1D). We found that immunization of NOD mice with insulin B-chain peptide B:9-23 followed by 72 h in vitro culture with B:9-23 peptide induces generation of CD4+CD25+ regulatory T cells. Route of immunization has a critical role in the generation of these cells. Non-autoimmune mice BALB/c, C57BL/6 and NOR did not show up regulation of CD4+CD25+ regulatory T cells. These cells secreted large amounts of TGF-beta and TNF-alpha with little or no IFN-gamma and IL-10. Adoptive transfer of these CD4+CD25+ regulatory T cells into NOD-SCID mice completely prevented the adoptive transfer of disease by diabetogenic T cells. Although, non-self antigenic OVA (323-339) peptide immunization and in vitro culture with OVA (323-339) peptide does result in up regulation of CD4+CD25+ T cells, these cells did not prevent transfer of diabetes. Our study for the first time identified the generation of antigen-specific CD4+CD25+ regulatory T cells specifically in response to immunization with B:9-23 peptide in NOD mice that are capable of blocking adoptive transfer of diabetes. Our results suggest the possibility of using autoantigens to induce antigen-specific regulatory T cells to prevent and regulate autoimmune diabetes.

Adoptive Transfer↗

Therapeutic efficacy of adoptive immunotherapy is predicated on in vivo antigen-specific proliferation of donor T cells.

Activated T cells with down-regulated L-selectin expression (L-sel(-)) from tumor-draining lymph nodes represent a potent source of specific immune effectors in adoptive immunotherapy. Using congenic pairs of mice and carboxyfluorescein diacetate succinimidyl ester-labeled L-sel(-) T cells, the current study analyzed in vivo proliferation of transferred cells. In the lung of MCA205 tumor-bearing mice, 6% or 0.3 x 10(6) of the 5 x 10(6) donor cells were identified 24 h after transfer. Vigorous proliferation of donor cells was evident on day 2, reaching a maximum on day 6. The proliferation was tumor-specific and CD4 T cells divided with greater magnitude than CD8 cells. Successful adoptive immunotherapy also required sublethal whole-body irradiation (WBI) of the recipient. WBI exerted its effects on facilitating specific T cell proliferation at the tumor site. Taken together, our results demonstrate that adoptively transferred T cells undergo extensive proliferation in response to the tumor and this response is associated with therapeutic efficacy.

Animals↗

Salivary cortisol levels in children adopted from romanian orphanages.

Six and a half years after adoption. 6- to 12-year-old children reared in Romanian orphanages for more than 8 months in their first years of life (RO. n = 18) had higher cortisol levels over the daytime hours than did early adopted (EA, < or = 4 months of age, n = 15) and Canadian born (CB, n = 27) children. The effect was marked, with 22% of the RO children exhibiting cortisol levels averaged over the day that exceeded the mean plus 2 SD of the EA and CB levels. Furthermore, the longer beyond 8 months that the RO children remained institutionalized the higher their cortisol levels. Cortisol levels for EA children did not differ in any respect from those of CB comparison children. This latter finding reduces but does not eliminate concerns that the results could be due to prenatal effects or birth family characteristics associated with orphanage placement. Neither age at cortisol sampling nor low IQ measured earlier appeared to explain the findings. Because the conditions in Romanian orphanages at the time these children were adopted were characterized by multiple risk factors, including gross privation of basic needs and exposure to infectious agents, the factor(s) that produced the increase in cortisol production cannot be determined. Nor could we determine whether these results reflected effects on the limbic-hypothalamic-pituitary-adrenal axis directly or were mediated by differences in parent-child interactions or family stress occasion by behavioral problems associated with prolonged orphanage care in this sample.

Adoption↗

Stoolmiller on restriction of range in adoption studies: a comment.

Stoolmiller has recently presented a model featuring an inferred dimension of family quality which, he suggests, is severely truncated in adoption studies such as the Texas Adoption Project, resulting in gross underestimation of the effects of shared family environment. We discuss some potential limitations of his approach in general and as he applies it to the Texas adoption data on IQ.

Adoption↗

Parental education and child's verbal IQ in adoptive and biological families in the National Longitudinal Study of Adolescent Health.

This study compared adoptive children and matched, biological children to estimate the genetic and environmental effect of years of mothers' and fathers' education on children's verbal intelligence (VIQ), as assessed by knowledge of vocabulary words. Adoptive and biological adolescent children in the National Longitudinal Study of Adolescent Health (Add Health) were matched for sex, age, parental education, and ethnicity. The adolescents all resided with two parents. Structural equation modeling was employed using Mx to estimate the genetic and transmissible environmental components of the correlation between parental education and children's VIQ. The mother-child and father-child correlations in biological families were .41 and .36, respectively, vs .16 and .18 in adoptive families. As suggested by these correlations, both genetic and shared environmental influences were statistically significant in the Mx models. We conclude that parental education exerts a modest shared environmental effect, explaining no more than 3 to 4% of the variation in verbal intelligence.

Adolescent↗

Finnish adoptive family study: sample selection and adoptee DSM-III-R diagnoses.

OBJECTIVE: To evaluate the genetic contribution to schizophrenia using an adoption design that disentangles genetic and environmental factors. METHOD: Finnish hospital diagnoses of schizophrenic/paranoid psychosis in a nationwide sample of adopting-away women are compared with DSM-III-R research diagnoses for these mothers. DSM-III-R diagnoses of their index offspring are blindly compared with adopted-away offspring of epidemiologically unscreened control mothers. RESULTS: Primary sampling diagnoses of index mothers were confirmed using DSM-III-R criteria. Lifetime prevalence of typical schizophrenia in 164 index adoptees was 6.7% (age-corrected morbid risk 8.1%), significantly different from 2.0% prevalence (2.3% age-corrected morbid risk) in 197 control adoptees. When adoptees with diagnoses of schizoaffective disorder, schizophreniform disorder, schizotypal disorder and affective psychoses were added, the contrast between the index and control adoptees increased. CONCLUSION: The genetic liability to 'typical' DSM-III-R schizophrenia is decisively confirmed. Additionally, the liability also extends to a broad spectrum of other psychotic and non-psychotic disorders.

Adoption↗

Maternal sensitivity, infant attachment, and temperament in early childhood predict adjustment in middle childhood: the case of adopted children and their biologically unrelated parents.

In a longitudinal study, internationally adopted children (N = 146) placed before 6 months of age were followed from infancy to age 7. Results showed that girls were better adjusted than boys, except in cognitive development, and that easy temperament was associated with higher levels of social, cognitive, and personality development and fewer behavior problems. Higher quality of child-mother relationships, in terms of attachment security and maternal sensitivity, uniquely predicted better social and cognitive development. The combination of attachment disorganization and difficult temperament predicted less optimal ego-control and lower levels of cognitive development. It is concluded that even in adopted children, who are not biologically related to their adoptive parents, early mother-infant interactions and attachment relationships predict later socioemotional and cognitive development, beyond infant temperament and gender.

Adoption↗

Genetic and environmental influences on MMPI factor scales: joint model fitting to twin and adoption data.

In general, the shared family environment appears to play a negligible role in determining individual differences in personality and interests. Nevertheless, scattered reports of significant shared environmental influence on such variables appear in the literature. Using data from the Texas Adoption Project (TAP), the current study attempted to replicate twin study findings of significant shared environmental variance on four of nine Minnesota Multiphasic Personality Inventory (MMPI) factor scales (Rose, 1988). Conventional behavioral genetic analyses of the adoption data agreed in affirming a significant shared environmental influence on individual differences in Religious Orthodoxy only. Subsequent simultaneous modeling of Rose's twin data and TAP adoption data resulted in three scales (Extraversion, Inadequacy, and Religious Orthodoxy) showing significant shared environmental influence. Again, effects were most substantial for Religious Orthodoxy, where the shared environment accounted for nearly 50% of the variance. It is argued that assortative mating cannot explain this finding.

Adolescent↗