Marathon runners (N = 22) who had completed at least seven marathons (X ± SEM = 23.6 ± 5.7) and had been training for marathon race events for at least 4 yr (12.3 ± 1.3) were compared with sedentary controls (N = 18). Although the two groups were of similar age (38.7 ± 1.5 and 43.9 ± 2.2 yr, respectively) and height, the marathon runners were significantly leaner and possessed a VO2max 60% higher than that of the controls. Neutrophil counts tended to be lower in the group of marathoners, while other leukocyte and lymphocyte subsets were similar to controls. Mitogen-induced lymphocyte proliferation did not differ between groups. Natural killer cell cyto-toxic activity (NKCA) was significantly higher in the marathoners versus controls (373 ± 38 vs 237 ± 41 total lytic units, respectively, a 57% difference, P = 0.02). For all subjects combined (N = 40) and within the group of marathon runners (N — 22), percent body fat was negatively correlated with NKCA (r = -0.48, P = 0.002; r = -0.49, P = 0.019, respectively), and age was negatively correlated with Con A-induccd lymphocyte proliferation (r = -0.41, P = 0.009; r = -0.53, P = 0.011, respectively). These data indicate that NKCA but not mitogen-induced lymphocyte proliferation is higher in marathon runners relative to sedentary controls.

L’idée qu’une cellule puisse effectuer la cytolyse de cellules transformées, comme une cellule Natural Killer (NK), tout en ayant la capacité de présenter des antigènes, comme une cellule dendritique (DC), peut sembler fantaisiste. Cependant, de telles cellules furent bel et bien identifiées chez la souris en 2006. Ces cellules, nommées Interferon-producing Killer Dendritic Cells (IKDC), furent l’objet d’une caractérisation extensive qui révéla leur énorme potentiel immunologique. La combinaison de fonctions associées à des cellules NK et à des DC a doté les IKDC d’un pouvoir antitumoral remarquable. D’ailleurs, il a été démontré que les IKDC sont plus efficaces que les cellules NK pour limiter la croissance tumorale. Ainsi, suite à leur découverte, les IKDC ont suscité beaucoup d’intérêt. Cependant, une controverse émergea sur la nature des IKDC. Plusieurs groupes indépendants tentèrent de reproduire les expériences attestant les fonctions de DC des IKDC, sans y parvenir. De plus, des études additionnelles révélèrent que les IKDC possèdent des similitudes très importantes avec les cellules NK. Ces observations ont mené la communauté scientifique à suggérer que les IKDC sont des cellules NK en état d’activation (aNK). Malgré cette controverse, les caractéristiques antitumorales des IKDC sont si uniques et considérables qu’il est primordial de poursuivre l’étude de ces cellules. Pour y arriver, il est essentiel de déterminer la nature des IKDC et de mettre fin à ce débat. Par la suite, il sera important d’identifier des façons de cibler spécifiquement les IKDC pour permettre leur usage dans le cadre de thérapies antitumorales. Ainsi, l’objectif de cette thèse est de définir l’identité des IKDC, puis de déterminer les facteurs génétiques responsables de la régulation de ces cellules. Nous avons démontré que les IKDC ne sont pas des cellules aNK, contrairement à ce qui avait été suggéré. Nous avons constaté que les IKDC prolifèrent activement et possèdent un phénotype unique, des caractéristiques associées à des cellules NK très immatures. Afin de déterminer si les IKDC peuvent acquérir un phénotype mature, nous avons effectué des expériences de transfert adoptif. Suite à leur injection in vivo, les IKDC acquièrent un phénotype de cellules matures, mais étonnamment, elles se différencient aussi en cellules NK. Ainsi, nous avons révélé que les IKDC sont un intermédiaire dans la différenciation des cellules NK. En parallèle, nous avons démontré que la proportion d’IKDC varie grandement entre des souris de fond génétique différent, indiquant que des facteurs génétiques sont impliqués dans la régulation de ces cellules. Nous avons alors effectué une analyse génétique qui a révélé que les IKDC sont régulées par des facteurs génétiques compris dans une région distale du chromosome 7. Les résultats présentés dans cette thèse constituent une avancée importante pour la recherche sur les IKDC. Ils ont permis de définir la nature des IKDC et d’identifier un intervalle génétique impliqué dans la régulation de ces cellules. Ces découvertes sont des connaissances précieuses pour l’identification des IKDC chez l’Homme et la création de nouvelles thérapies dans la lutte contre le cancer. / The idea that a cell could kill transformed cells, like a Natural Killer (NK) cell, all the while exhibiting also the capacity to present antigens to T cells, like a Dendritic Cell (DC), may seem farfetched. However, in mice, a cell presenting these specific properties was identified in 2006. These cells were named Interferon-producing Killer Dendritic Cells (IKDC) and extensive studies revealed that they were endowed with an important immunological potential. Indeed, the fact that IKDCs exhibit properties of both DC and NK cells conferred them with an exceptional anti-tumor potential. Notably, on a per cell basis, the in vivo anti-tumor activity of IKDCs is more efficient than NK cells. Therefore, following their identification, IKDCs showed great therapeutic promise. However, a debate on the cell lineage origin of IKDCs emerged. Several independent groups could not replicate the finding that IKDCs showed functional antigen-presentation properties similar to DCs. Also, additional studies revealed that IKDCs are very similar to NK cells. These and other observations led the scientific community to believe that IKDCs were activated NK cells. Despite this controversy, IKDCs clearly exhibit a unique and outstanding anti-tumor potential, highlighting the relevance to further explore these cells. We must first close the debate regarding the lineage origin of IKDCs. We subsequently need to identify a means to specifically target IKDCs to facilitate their use in novel anti-tumor therapies. Thus, the objective of my thesis is first, to define the identity of IKDCs and second, to determine the genetic factors implicated in the regulation of these cells. For the first objective, we demonstrated that IKDCs do not represent activated NK cells, as previously suggested. We show that IKDCs are highly proliferative and exhibit a unique phenotype associated with very immature NK cells. In an attempt to verify if IKDCs could acquire a mature phenotype, we conducted an adoptive transfer experiment. We found that, after adoptive transfer, IKDCs adopt a mature phenotype, but also surprisingly differentiate into NK cells. These findings indicate that IKDCs represent an intermediate in NK-cell differentiation. For the second objective, we demonstrated that the IKDC proportion was highly variable between strains of different background origins, indicating that these cells are regulated by genetic factors. A genetic study revealed that genetic factors in distal arm of chromosome 7 associate with the proportion of IKDCs. The results presented in this thesis represent an important breakthrough for the research on IKDCs. They allowed to define the cell lineage origin of IKDCs and to identify a genetic region involved in the regulation of this cell type. These discoveries are valuable knowledge for the identification of human IKDCs and the development of novel anti-tumor therapies.

L’idée qu’une cellule puisse effectuer la cytolyse de cellules transformées, comme une cellule Natural Killer (NK), tout en ayant la capacité de présenter des antigènes, comme une cellule dendritique (DC), peut sembler fantaisiste. Cependant, de telles cellules furent bel et bien identifiées chez la souris en 2006. Ces cellules, nommées Interferon-producing Killer Dendritic Cells (IKDC), furent l’objet d’une caractérisation extensive qui révéla leur énorme potentiel immunologique. La combinaison de fonctions associées à des cellules NK et à des DC a doté les IKDC d’un pouvoir antitumoral remarquable. D’ailleurs, il a été démontré que les IKDC sont plus efficaces que les cellules NK pour limiter la croissance tumorale. Ainsi, suite à leur découverte, les IKDC ont suscité beaucoup d’intérêt. Cependant, une controverse émergea sur la nature des IKDC. Plusieurs groupes indépendants tentèrent de reproduire les expériences attestant les fonctions de DC des IKDC, sans y parvenir. De plus, des études additionnelles révélèrent que les IKDC possèdent des similitudes très importantes avec les cellules NK. Ces observations ont mené la communauté scientifique à suggérer que les IKDC sont des cellules NK en état d’activation (aNK). Malgré cette controverse, les caractéristiques antitumorales des IKDC sont si uniques et considérables qu’il est primordial de poursuivre l’étude de ces cellules. Pour y arriver, il est essentiel de déterminer la nature des IKDC et de mettre fin à ce débat. Par la suite, il sera important d’identifier des façons de cibler spécifiquement les IKDC pour permettre leur usage dans le cadre de thérapies antitumorales. Ainsi, l’objectif de cette thèse est de définir l’identité des IKDC, puis de déterminer les facteurs génétiques responsables de la régulation de ces cellules. Nous avons démontré que les IKDC ne sont pas des cellules aNK, contrairement à ce qui avait été suggéré. Nous avons constaté que les IKDC prolifèrent activement et possèdent un phénotype unique, des caractéristiques associées à des cellules NK très immatures. Afin de déterminer si les IKDC peuvent acquérir un phénotype mature, nous avons effectué des expériences de transfert adoptif. Suite à leur injection in vivo, les IKDC acquièrent un phénotype de cellules matures, mais étonnamment, elles se différencient aussi en cellules NK. Ainsi, nous avons révélé que les IKDC sont un intermédiaire dans la différenciation des cellules NK. En parallèle, nous avons démontré que la proportion d’IKDC varie grandement entre des souris de fond génétique différent, indiquant que des facteurs génétiques sont impliqués dans la régulation de ces cellules. Nous avons alors effectué une analyse génétique qui a révélé que les IKDC sont régulées par des facteurs génétiques compris dans une région distale du chromosome 7. Les résultats présentés dans cette thèse constituent une avancée importante pour la recherche sur les IKDC. Ils ont permis de définir la nature des IKDC et d’identifier un intervalle génétique impliqué dans la régulation de ces cellules. Ces découvertes sont des connaissances précieuses pour l’identification des IKDC chez l’Homme et la création de nouvelles thérapies dans la lutte contre le cancer. / The idea that a cell could kill transformed cells, like a Natural Killer (NK) cell, all the while exhibiting also the capacity to present antigens to T cells, like a Dendritic Cell (DC), may seem farfetched. However, in mice, a cell presenting these specific properties was identified in 2006. These cells were named Interferon-producing Killer Dendritic Cells (IKDC) and extensive studies revealed that they were endowed with an important immunological potential. Indeed, the fact that IKDCs exhibit properties of both DC and NK cells conferred them with an exceptional anti-tumor potential. Notably, on a per cell basis, the in vivo anti-tumor activity of IKDCs is more efficient than NK cells. Therefore, following their identification, IKDCs showed great therapeutic promise. However, a debate on the cell lineage origin of IKDCs emerged. Several independent groups could not replicate the finding that IKDCs showed functional antigen-presentation properties similar to DCs. Also, additional studies revealed that IKDCs are very similar to NK cells. These and other observations led the scientific community to believe that IKDCs were activated NK cells. Despite this controversy, IKDCs clearly exhibit a unique and outstanding anti-tumor potential, highlighting the relevance to further explore these cells. We must first close the debate regarding the lineage origin of IKDCs. We subsequently need to identify a means to specifically target IKDCs to facilitate their use in novel anti-tumor therapies. Thus, the objective of my thesis is first, to define the identity of IKDCs and second, to determine the genetic factors implicated in the regulation of these cells. For the first objective, we demonstrated that IKDCs do not represent activated NK cells, as previously suggested. We show that IKDCs are highly proliferative and exhibit a unique phenotype associated with very immature NK cells. In an attempt to verify if IKDCs could acquire a mature phenotype, we conducted an adoptive transfer experiment. We found that, after adoptive transfer, IKDCs adopt a mature phenotype, but also surprisingly differentiate into NK cells. These findings indicate that IKDCs represent an intermediate in NK-cell differentiation. For the second objective, we demonstrated that the IKDC proportion was highly variable between strains of different background origins, indicating that these cells are regulated by genetic factors. A genetic study revealed that genetic factors in distal arm of chromosome 7 associate with the proportion of IKDCs. The results presented in this thesis represent an important breakthrough for the research on IKDCs. They allowed to define the cell lineage origin of IKDCs and to identify a genetic region involved in the regulation of this cell type. These discoveries are valuable knowledge for the identification of human IKDCs and the development of novel anti-tumor therapies.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Thesis (PhD)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: In the current study, two isolates showing killer activity against several wine yeast species in a previous study were identified to strain level and found to belong to the yeast species Candida pyralidae. The identified yeast strains and a Kluyveromyces wickerhamii yeast strain used as a control exhibited killer activity against B. bruxellensis known for its spoilage characteristics in red wine, and against several strains of the genus Brettanomyces on white and red grape juice medium. The killer yeasts inhibited neither the growth of S. cerevisiae nor that of the lactic acid bacteria Oenococcus oeni and Lactobacillus plantarum strains. Yeasts are reported to secrete killer toxins, which can play a role in yeast microbial interactions under winemaking conditions. The C. pyralidae strains were found to secrete two novel killer toxins, designated CpKT1 and CpKT2. These killer toxins were stable and active under winemaking conditions, pH 3.5 - 4.5 and temperature ranges between 15 and 25°C. Ethanol and sugar concentrations found during winemaking did not affect the activity and stability of these killer toxins. Although, the killer toxins differed with regards to their biochemical and environmental stability and activity, they were found to have a similar mode of action. The killer toxins induced a fungistatic effect on B. bruxellensis sensitive cells in addition to binding to the cell wall of the sensitive cells, inducing cell surface and plasma membrane damage as did the Kwkt killer toxin secreted by K. wickerhamii. According to the author’s knowledge this is the first report on the identification of novel killer toxins secreted by C. pyralidae strains isolated from a wine environment as well as the identification of the mode of action of killer toxins on B. bruxellensis cells. This indeed provides great research scope in this field. The exoproteomes consisting of the killer toxins Kwkt, CpKT1 and CpKT2 revealed the presence of exo-glucanases and glucosidases, respectively. The enzymes KwExg1 (exoglucanase) and KwSun4 (glucosidase) retrieved from K. wickerhamii’s exoproteome were identified as the potential toxins, but their killer activity could not be confirmed. These findings suggest that hydrolytic enzymes possess killer activity, as previously reported in literature. However, further investigation is needed to identify the killer toxins characterized in this study. / AFRIKAANSE OPSOMMING: In die huidige studie is twee isolate wat in ’n vorige studie “killer” aktiwiteit teenoor verskeie wyngisspesies vertoon het, tot op rasvlak geïdentifiseer en daar is gevind dat hulle aan die gisspesie Candida pyralidae behoort. Die geïdentifiseerde gisrasse en ’n Kluyveromyces wickerhamii gisras wat as kontrole gebruik is, het “killer” aktiwiteit getoon teenoor B. bruxellensis, wat bekend is vir sy bederfkarakter in rooi wyn, en ook teenoor verskeie rasse van die genus Brettanomyces in wit en rooi druiwesapmedium. Die “killer” giste het nie die groei van óf S. cerevisiae óf van die melksuurbakteria Oenococcus oeni en Lactobacillus plantarum-rasse geïnhibeer nie. Giste word berig om “killer” gifstowwe uit te skei, wat ’n rol kan speel in gis mikrobiese interaksies onder wynbereidingstoestande. Die C. pyralidae-rasse is gevind om twee nuwe “killer” gifstowwe af te skei, wat CpKT1 en CpKT2 genoem is. Hierdie “killer” gifstowwe was stabiel en aktief onder wynbereidingstoestande, pH 3.5 - 4.5 en temperatuur tussen 15 en 25°C. Die etanol- en suikerkonsentrasies wat onder wynbereiding voorkom, het nie die aktiwiteit en stabiliteit van hierdie “killer” gifstowwe beïnvloed nie. Hoewel die “killer” gifstowwe met betrekking tot hulle biochemiese en omgewingstabiliteit en aktiwiteit verskil het, is daar gevind dat hulle ’n eenderse modus van aksie het. Die “killer” gifstowwe het ’n fungistatiese effek op B. bruxellensis sensitiewe selle geïnduseer, buiten dat dit aan die selwand van die sensitiewe selle gebind het, en het seloppervlak- en plasma-membraanskade geïnduseer, net soos die Kwkt “killer” gifstof wat deur K. wickerhamii afgeskei is. So ver die skrywer weet, is hierdie die eerste verslag van die identifisering van nuwe “killer” gifstowwe wat deur C. pyralidae rasse afgeskei word wat uit ’n wynomgewing geïsoleer is, asook van die identifikasie van die modus van aksie van “killer” gifstof op B. bruxellensis selle. Dit verbreed dus beslis die navorsingsomvang van hierdie gebied. Die eksoproteome, bestaande uit die “killer” gifstowwe Kwkt, CpKT1 en CpKT2, het die teenwoordigheid van ekso-glukanases en glukosidases onderskeidelik onthul. Die ensieme KwExg1 (eksoglukanase) en KwSun4 (glukosidase) wat vanuit K. wickerhamii se eksoproteoom herwin is, is as die potensiële gifstowwe geïdentifiseer, maar hulle “killer” aktiwiteit kon nie bevestig word nie. Hierdie bevindings suggereer dat hidrolitiese ensieme “killer” aktiwiteit besit, soos voorheen in die literatuur berig is. Verdere ondersoeke word egter benodig om die “killer” gifstowwe wat in hierdie studie gekarakteriseer is, te identifiseer.

No description available.

Natural killer (NK) cells are lymphocytes of the innate immune system that play a major role in the destruction of both tumours and virally-infected cells. The cytotoxicity of NK cells is tightly controlled by signals received through activating and inhibitory receptors. NK cells express a variety of inhibitory receptors such as Ly49 receptors. Ly49 receptors bind to class I MHC molecules that expressed on normal cells. Using Ly49-deficient (NKCKD) mice we show that Ly49-KD NK cells successfully recognize and kill influenza virus-infected cells and that NKCKD mice exhibit better survival than wild-type mice. Moreover, influenza virus infection has a propensity to upregulate cell surface expression of MHC-I on murine lung epithelial cells in vivo. Significantly, we demonstrate increased lung damage of WT-mice versus NKCKD mice after influenza virus infection as determined by histological analyses. This data indicated that absence of Ly49 inhibitory NK receptors greatly enhances survival of infected mice.

Influenza A virus (IAV) infection is a highly contagious respiratory disease, which can cause substantial morbidity and occasionally death. Invariant natural killer T (iNKT) cells, a subset of CD1d-restricted T lymphocytes, have been identified as important modulators of immunity, mediating both pro- and anti-inflammatory responses. We show that iNKTs play an important role for the generation of protective innate and adaptive immune responses to IAV, and enhance heterotypic immunity to influenza virus following vaccination with a novel pseudotyped virus, S-FLU, which lacks HA expression. iNKT-deficient mice (Jα18^-/-) showed increased susceptibility and lung pathology following IAV infection, which correlated with an exaggerated accumulation of inflammatory monocytes and neutrophils in the lung. Consistent with these findings, we demonstrated in IAV-infected CD1d^-/-:CD1d^{+/+} mixed bone marrow chimeric mice, that the lack of CD1d expression on myeloid cells purified from the lungs of IAV-infected mice significantly increased expression of genes linked to cell activation, survival and polarisation between pro- and antiinflammatory responses. We extended these results by examining the role of chemokine signalling during IAV infection, and identified a novel role for fractalkine (CX3CL1) and its receptor (CX3CR1) in innate immune cell recruitment. The use of CX3CR1-iNKT cell double knockout mice revealed that, although upregulated in Jα18^-/- mice, CX3CR1-CX3CL1 signalling is not required for cell migration during exacerbated IAV-responses. Finally, we showed that iNKT-deficient mice displayed reduced longevity of peripheral IAVspecific CD8^+ T cells following S-FLU vaccination, compared with wild-type mice. S-FLU vaccination protected mice following 5 day heterotypic challenge, however vaccinated mice exhibited reduced viral clearance, and importantly a significant reduction in IAV specific memory T cell response, suggesting a possible role of iNKT cells during T cell priming in modulating the lifespan of antigen-specific T cell responses. Although additional experiments are warranted, these results suggest that harnessing iNKT cells should be considered to modulate the innate and adaptive immune response to optimise heterotypic vaccine design and for therapeutic intervention against acute influenza infection.

Danazol, um andrógeno sintético, desenvolvido em 1970 para o tratamento da endometriose, tem aplicação controvérsia no pré-tratamento de mulheres com infertilidade sem causa aparente, abortos recorrentes e que seriam submetidas à fertilização com transferência de embrião. O conhecimento sobre a atuação deste medicamento sobre a incidência de células uNK, morfologia uterina e funções cognitivas após sua administração ainda são muito fragmentados e não conclusivos. Os objetivos deste trabalho foram avaliar os efeitos das diferentes concentrações do danazol no comportamento de camundongos prenhes, bem como, as possíveis alterações na implantação, viabilidade, perda embrionára, morfologia do útero e incidência dos subtipos de células uNK reativos para lectina DBA. Foram utilizadas noventa e três fêmeas da linhagem Swiss, obtidas do Biotério da Universidade Federal de Alfenas, tratadas durante 14 dias por meio de gavagem com água destilada + tween 1% (grupo controle) e com água destilada + tween 1% + danazol nas concentrações de 0,75; 7,5; ou 75 mg/kg. O tratamento nos animais foi suspenso por 4 dias e após este prazo as fêmeas foram acasaladas com os machos. O dia em que foi constatada a presença do tampão vaginal foi considerado o 1° dia de gestação. Os testes comportamentais foram realizados e os animais foram sacrificados no 6º, 8º, 10º, 12º e 15º dia de gestação (ddg). Os resultados obtidos mostraram que danazol na dosagem de 0,75 mg/Kg apresentou efeito ansiogênico em camundongos e diminuiu a retenção de memória de curto prazo no 10º ddg. Na concentração de 7,5 mg/Kg provocou aumento da retenção de memória a curto prazo e estado equivalente ao depressivo. A diminuição do tempo de exploração global, à medida que a concentração do hormônio foi aumentada sugere efeito dose dependente de danazol no estabelecimento de estado de sonolência e apatia em camundongos no 10º ddg. Danazol provocou atraso no período para sucesso da cópula, no entanto, aumentou a taxa de prenhez sem reduzir o número de corpus lúteos em todas as dosagens utilizadas. Nos camundongos tratados com a dosagem 7,5 mg/Kg de danazol foram observadas, a partir do 10º ddg, muitas células grandes e de morfologia equivalente às células trofoblásticas gigantes ao redor dos vasos que nutrem a decídua e o embrião, o que sugere que está droga aumenta a invasividade do trofoblasto a partir do período médio da prenhez. Danazol na concentração de 7,5 mg/Kg provocou aumento no número de células uNK, enquanto dosagens menores (0,75 mg/Kg) e maiores (75 mg/Kg) causaram diminuição neste número, demonstrando o efeito dose dependente de danazol no número de células uNK. A análise dos diferentes subtipos de uNK no útero mostrou que danazol aumenta a migração destas células, podendo prejudicar a proliferação celular em doses elevadas e acelerar a diferenciação celular em áreas próximas ao embrião. As maiores alterações relacionadas à ansiedade e memória observadas no 10º ddg, associadas às alterações morfológicas nos sítios de implantação embrionária e aumento na proliferação de células natural killer uterinas também neste período, demonstram a integração entre sistemas nervoso, endócrino e imunológico durante a prenhez e a ação dose-dependente de danazol modulando as respostas destes sistemas. / Danazol, a synthetic androgen, developed in 1970 for the treatment of endometriosis, has a controversial application on the pre-treatment of women with unexplained infertility, recurrent miscarriage, and the ones that would undergo fertilization with embryo transfer. The knowledge about the way this compound works after its administration on the incidence of uNK cells, uterine morphology and cognitive functions is still incomplete and inconclusive. Our objectives were to evaluate the effects of different concentrations of danazol on the behavior of pregnant mice, as well as possible changes in implantation, viability, embryonic loss, morphology of the uterus and the incidence of DBA lectin-reactive uNK cell subtype. Ninety-three Swiss female mice from Universidade Federal de Alfenas vivarium were treated for 14 days by gavage with either distilled water + 1% Tween (control) or distilled water + 1% Tween + 0.75, 7.5 or 75 mg/kg of danazol. Females were mated with males after suspension of treatment during four days. Day 1 of gestation was established as the day when the presence of vaginal plug was detected. Behavioral tests were performed and the animals were sacrificed at 6th, 8th, 10th, 12th and 15th gestation days (gd). The data showed that 0.75 mg/kg of danazol caused an anxiogenic effect on mice and decreased the retention of short-term memory in the 10th gd. 7.5 mg/kg of this drug increased the retention of short-term memory and developed a depression-like state on the animals. The fact that gradual increase on hormone concentration caused gradual decrease on the global exploration time suggests a dose-dependent effect of danazol on establishing a state of somnolence and apathy in mice in the 10th ddg. Danazol caused a delay on time of successful mating, however, it increased the rate of pregnancy without reducing the number of corpus luteum in all dosages used. In mice treated with 7.5 mg/kg of danazol, many large cells with a trophoblast-like morphology were observed around vessels that nourish the decidua and the embryo starting at 10th gd, what suggests that this drug increases the invasiveness of the trophoblast starting in the middle period of pregnancy. 7.5 mg/kg of danazol increased the number of uNK cells, whereas the lowest (0.75 mg/ kg) and the highest (75 mg / kg) doses caused a decrease in this number, demonstrating a dose dependent effect of danazol on the number of uNK cells. The analysis of different subtypes of uNK cells in the uterus showed that danazol increases the migration of these cells and can also impair cell proliferation on higher doses. It also suggests that danazol can stimulate cell differentiation in areas close to the embryo. The major changes related to anxiety and memory observed in the 10th gd, associated with morphological changes in the sites of embryo implantation and increase in proliferation of uterine natural killer cells in this period, demonstrate the integration between the nervous, endocrine and immune systems during pregnancy and the dose-dependent action of danazol in the modulation of these systems. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES