[Effect of transfusing blood components containing Babesia microti on B. microti infection in BALB/c mice].

OBJECTIVE: To evaluate the effects of intravenous injection of different blood components containing Babesia microti on B. microti infection in mice. METHODS: Healthy mice were infected with B. microti, and then blood samples were collected from the mouse orbit to prepare whole blood, serum-free blood components and pure red blood cells containing B. microti. Twenty seven BALB/c mice were divided into three groups, including the whole blood group, the serum-free blood component group and the pure red blood cell group, of 9 mice in each group, and then, each group was divided into three subgroups, of 3 mice in each subgroup, which were injected with 100 µL of blood components containing B. microti at concentrations of 9.00, 0.90, 0.09 B. microti parasites/µL (900, 90, 9 B. microti parasites) via the tail vein, respectively. Blood samples were collected from the mouse tail tip every other day since one day post-injection to prepare thin blood smears. Following Giemsa staining of blood smears, B. microti infection was identified in red blood cells using microscopy. RESULTS: Following injection of 900 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group and the serum-free blood component group 3 days post-injection, and the density of B. microti parasites started to increase 15 days post-injection and peaked 21 days post-injection, with 2.21% and 1.76% rates of B. microti infection in red blood cells, respectively. Subsequently, the density of B. microti parasites declined, and the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was found in the peripheral blood in the pure red blood cell group. Following injection of 90 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group 3 days post-injection, and the density of B. microti parasites increased 15 days post-injection and peaked 21 days post-injection, with a 1.35% rate of B. microti infection in red blood cells, while the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was detected in the peripheral blood in the serum-free blood component group or the pure red blood cell group. Following injection of 9 B. microti parasites, no B. microti was detected in the peripheral blood in the whole blood group, the serum-free blood component group or the pure red blood cell group. CONCLUSIONS: Blood components and dose of B. microti parasites may affect intravenous injection of B. microti injection in mice, and transfusion of blood components may case a risk of Babesia infection.

Effect of transfusing blood components containing Babesia microti on B. microti infection in BALB/c mice / 中国血吸虫病防治杂志

Objective To evaluate the effects of intravenous injection of different blood components containing Babesia microti on B. microti infection in mice. Methods Healthy mice were infected with B. microti, and then blood samples were collected from the mouse orbit to prepare whole blood, serum-free blood components and pure red blood cells containing B. microti. Twenty seven BALB/c mice were divided into three groups, including the whole blood group, the serum-free blood component group and the pure red blood cell group, of 9 mice in each group, and then, each group was divided into three subgroups, of 3 mice in each subgroup, which were injected with 100 μL of blood components containing B. microti at concentrations of 9.00, 0.90, 0.09 B. microti parasites/μL (900, 90, 9 B. microti parasites) via the tail vein, respectively. Blood samples were collected from the mouse tail tip every other day since one day post-injection to prepare thin blood smears. Following Giemsa staining of blood smears, B. microti infection was identified in red blood cells using microscopy. Results Following injection of 900 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group and the serum-free blood component group 3 days post-injection, and the density of B. microti parasites started to increase 15 days post-injection and peaked 21 days post-injection, with 2.21% and 1.76% rates of B. microti infection in red blood cells, respectively. Subsequently, the density of B. microti parasites declined, and the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was found in the peripheral blood in the pure red blood cell group. Following injection of 90 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group 3 days post-injection, and the density of B. microti parasites increased 15 days post-injection and peaked 21 days post-injection, with a 1.35% rate of B. microti infection in red blood cells, while the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was detected in the peripheral blood in the serum-free blood component group or the pure red blood cell group. Following injection of 9 B. microti parasites, no B. microti was detected in the peripheral blood in the whole blood group, the serum-free blood component group or the pure red blood cell group. Conclusion Blood components and dose of B. microti parasites may affect intravenous injection of B. microti injection in mice, and transfusion of blood components may case a risk of Babesia infection.

Effect of transfusing blood components containing Babesia microti on B. microti infection in BALB/c mice / 中国血吸虫病防治杂志

Objective To evaluate the effects of intravenous injection of different blood components containing Babesia microti on B. microti infection in mice. Methods Healthy mice were infected with B. microti, and then blood samples were collected from the mouse orbit to prepare whole blood, serum-free blood components and pure red blood cells containing B. microti. Twenty seven BALB/c mice were divided into three groups, including the whole blood group, the serum-free blood component group and the pure red blood cell group, of 9 mice in each group, and then, each group was divided into three subgroups, of 3 mice in each subgroup, which were injected with 100 μL of blood components containing B. microti at concentrations of 9.00, 0.90, 0.09 B. microti parasites/μL (900, 90, 9 B. microti parasites) via the tail vein, respectively. Blood samples were collected from the mouse tail tip every other day since one day post-injection to prepare thin blood smears. Following Giemsa staining of blood smears, B. microti infection was identified in red blood cells using microscopy. Results Following injection of 900 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group and the serum-free blood component group 3 days post-injection, and the density of B. microti parasites started to increase 15 days post-injection and peaked 21 days post-injection, with 2.21% and 1.76% rates of B. microti infection in red blood cells, respectively. Subsequently, the density of B. microti parasites declined, and the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was found in the peripheral blood in the pure red blood cell group. Following injection of 90 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group 3 days post-injection, and the density of B. microti parasites increased 15 days post-injection and peaked 21 days post-injection, with a 1.35% rate of B. microti infection in red blood cells, while the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was detected in the peripheral blood in the serum-free blood component group or the pure red blood cell group. Following injection of 9 B. microti parasites, no B. microti was detected in the peripheral blood in the whole blood group, the serum-free blood component group or the pure red blood cell group. Conclusion Blood components and dose of B. microti parasites may affect intravenous injection of B. microti injection in mice, and transfusion of blood components may case a risk of Babesia infection.

Genetic diversity and phylogenetic analysis of EG95 sequences of Echinococcus granulosus: Implications for EG95 vaccine application

OBJECTIVE@#To analyse the genetic variability of EG95 sequences and provide guidance for EG95 vaccine application against Echinococcus granulosus (E. granulosus).@*METHODS@#We analysed EG95 polymorphism by collecting total 97 different E. granulosus isolates from 12 different host species that originated from 10 different countries. Multiple sequence alignments and the homology were performed by Lasergene 1 (DNASTAR Inc., Madison, WI), and the phylogenetic analysis was performed by using MEGA5.1 (CEMI, Tempe, AZ, USA). In addition, linear and conformational epitopes were analysed, including secondary structure, NXT/S glycosylation, fibronectin type III (FnIII) domain and glycosylphosphatidylinositol anchor signal (GPI-anchor). The secondary structure was predicted by PSIPRED method.@*RESULTS@#Our results indicated that most isolates overall shared 72.6-100% identity in EG95 gene sequence with the published standard EG95 sequence, X90928. However, EG95 gene indeed has polymorphism in different isolates. Phylogenetic analysis showed that different isolates could be divided into three subgroups. Subgroup 1 contained 87 isolates while Subgroup 2 and Subgroup 3 consisted of 3 and 7 isolates, respectively. Four sequences cloned from oncosphere shared a high identity with the parental sequence of the current vaccine, X90928, and they belonged to Subgroup 1. However, in comparison to X90928, several amino acid mutations occurred in most isolates besides oncosphere, which potentially altered the immunodominant linear epitopes, glycosylation sites and secondary structures in EG95 genes. All these variations might change their previous antigenicity and thereby affecting the efficacy of current EG95 vaccine.@*CONCLUSIONS@#This study reveals the genetic variability of EG95 sequences in different E. granulosus isolates, and proposed that more vaccination trials would be needed to test the effectiveness of current EG95 vaccine against distinct isolates in different countries.

Genetic diversity and phylogenetic analysis of EG95 sequences of Echinococcus granulosus: Implications for EG95 vaccine application

Objective To analyse the genetic variability of EG95 sequences and provide guidance for EG95 vaccine application against Echinococcus granulosus (E. granulosus). Methods We analysed EG95 polymorphism by collecting total 97 different E. granulosus isolates from 12 different host species that originated from 10 different countries. Multiple sequence alignments and the homology were performed by Lasergene 1 (DNASTAR Inc., Madison, WI), and the phylogenetic analysis was performed by using MEGA5.1 (CEMI, Tempe, AZ, USA). In addition, linear and conformational epitopes were analysed, including secondary structure, NXT/S glycosylation, fibronectin type III (FnIII) domain and glycosylphosphatidylinositol anchor signal (GPI-anchor). The secondary structure was predicted by PSIPRED method. Results Our results indicated that most isolates overall shared 72.6–100% identity in EG95 gene sequence with the published standard EG95 sequence, X90928. However, EG95 gene indeed has polymorphism in different isolates. Phylogenetic analysis showed that different isolates could be divided into three subgroups. Subgroup 1 contained 87 isolates while Subgroup 2 and Subgroup 3 consisted of 3 and 7 isolates, respectively. Four sequences cloned from oncosphere shared a high identity with the parental sequence of the current vaccine, X90928, and they belonged to Subgroup 1. However, in comparison to X90928, several amino acid mutations occurred in most isolates besides oncosphere, which potentially altered the immunodominant linear epitopes, glycosylation sites and secondary structures in EG95 genes. All these variations might change their previous antigenicity and thereby affecting the efficacy of current EG95 vaccine. Conclusions This study reveals the genetic variability of EG95 sequences in different E. granulosus isolates, and proposed that more vaccination trials would be needed to test the effectiveness of current EG95 vaccine against distinct isolates in different countries.

[Splenic lymphocyte immune response induced by intranasal immunization with recombinant Toxoplasma gondii profilin in mice].

OBJECTIVE: To observe the splenocytes immune response elicited by different concentrations of recombinant Toxoplasma gondii profiling (rTgPRF) through the nasal route, and determine the optimal dose. METHODS: Fifty female BALB/c mice were randomly divided into 5 groups. The immunized groups were intranasally administered with 10, 20, 30 µg or 40 µg of rTgPRF that was separately dissolved in 20 µl of phosphate-buffered saline (PBS) on days 0, 14, and 21 respectively, while the control mice were given PBS solution instead. Two weeks after the last immunization, all mice were killed. Under asceptic conditions, the spleens from the immunized mice were dissected, and then the splenocyte proliferative responses in vitro were tested by CCK-8 kit. The levels of IFN-γ, IL-2, IL-4 and IL-10 of splenocyte culture supernatant were detected by ELISA. RESULTS: Compared to the control group, the splenocytes from the 30 µg and 40 µg groups exhibited a significantly higher proliferative response to rTgPRF (P < 0.05), and SI from the 30 µg rTgPRF group was higher than that from the 40 µg group (P < 0.05). The levels of IFN-γ in all the immunized groups (P < 0.05) and IL-2 in the 20, 30 µg and 40 µg groups were significantly stronger than those in the control (P < 0.05), and the 30 µg group presented the highest concentrations of IFN-γ (P < 0.01) and IL-2 (P < 0.01). There were no statistical differences among the groups in the levels of IL-4 and IL-10. CONCLUSIONS: The intranasal immunization with rTgPRF can induce the splenocyteproliferation and Th1-type mediated immunity. The best immunized dose is confirmed as 30 µg.

[Changes and significance of inflammatory cytokines in sera of mice chronically infected with larvae of Echinococcus granulosus].

OBJECTIVE: To investigate the levels of cytokines in the sera of mice chronically infected with the larvae of Echinococcus granulosus, and explore the mechanisms of immune regulation against parasite infection. METHODS: The protoscoleces were isolated from the livers and lungs of sheep infected with E. granulosus, and then inoculated intraperitoneally to BALB/c mice (2 000 for each mouse), to establish the mouse model of E. granulosus infection. The mice in the control group were injected intraperitoneally with an equal volume of PBS. The sera of both control and infected mice were collected to test the levels of multiple cytokines by using the Cytometric Bead Array (CBA) 5 months post-infection. RESULTS: In contrast to the control group, the multiple cysts were found in the abdominal cavity, livers and lungs of the infected mice. Moreover, the levels of inflammatory cytokines such as IL-17A, IL-6, IFN-γ, MCP-1, IL-12P70 and TNF-α in the sera of the infected mice were significantly higher than those in the control group (t = 2.713-9.255, all P < 0.05) while the levels of anti-inflammatory cytokine IL-10 were elevated post-infection (t = 3.936, P < 0.001). CONCLUSIONS: Higher inflammatory cytokines in the mice chronically infected with the larvae of E. granulosus, may benefit for the limitation of parasite growth.

Effects of p38MAPK inhibitor on the occurrence of acute GVHD and intestine damage after allogeneic hematopoietic stem cell transplantation in mice / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effects of p38MAPK inhibitor SB203580 (SB) on the occurrence of acute GVHD and intestine damage after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in mice.</p><p><b>METHODS</b>Sixty BALB/c mice, as recipients, were randomized to control group, irradiation group, model group and intervention group. C57BL/6 mice, as donors, were raised to prepare the bone marrow cells (BMCs) and spleen cells (SCs), which were injected into irradiated recipients mice by tail vein. Except control group, other groups accepted 7.5Gy total body irradiation. Model group and intervention group were infused with BMCs 5×10⁶ and SCs 5×10⁵ by less than 4 h after irradiation. SB was injected into intervention group by intraperitoneally, but only DMSO for model group. The general status and survival rate of each group were evaluated. The expression of p-p38MAPK, Fas and FasL in intestine were determined by RT-PCR, Western blot and immunohistochemistry (IHC).</p><p><b>RESULTS</b>The weight changes of intervention group (13.00±0.50)% was significantly lighter than that of model group (25.00±0.75)% (P<0.05). The clinical score of acute GVHD in the intervention group (3.33±0.82) was significantly lower than that of model group (6.33±1.36) (P<0.05). The expression levels of p-p38MAPK, Fas and FasL in small intestine of intervention group (1.43±0.02, 0.81±0.03, 0.97±0.03) were lower than those of model group (1.76±0.05, 1.52±0.04, 1.48±0.04).</p><p><b>CONCLUSION</b>SB inhibited the activation of p38MAPK and Fas/ FasL signal pathway and alleviated the apoptosis of small intestine. And SB could relieve small intestine damages induced by allogeneic T lymphocytes.</p>