Hematological, hemorheological, immunological, and morphological studies of spleen autotransplantation in mice: preliminary results

Using a spleen autotransplantation model, we conducted hematological, hemorheological, immunological, and morphological studies in mice 6 weeks after splenectomy. Sixty male and female A/J inbred mice were equally divided into 3 groups: 1) SE group, splenectomy was performed; 2) AU group, spleen chips were autotransplanted into the omentum without vascular anastomosis following splenectomy; and 3) C group (controls), no intervention in these mice. At postoperative week 6, the following studies were performed: 1) measurement of hematological parameters; 2) hemorheological studies, including relative cell transit time (RCTT) and fibrinogen levels; and 3) activity of peripheral phagocytes, measured by zymozan-induced chemiluminescence, which was calculated in stimulation index values (SI). In addition, histological investigations of autotransplants were conducted. Erythrocyte mean cell volume and platelet counts, RCTT, fibrinogen levels, and activity of phagocytes were significantly higher in the SE group, compared to those in the C group. In the AU group, these parameters were similar to those in the C group. Morphologically, the transplanted spleen showed normal histology. These data indicate that the transplanted spleens restored their function. We conclude that spleen autotransplantation reserves the normal morphology of spleen and restores most of the spleen's hematological, hemorheological, and immunological functions. Both SI index and erythrocyte deformability can be an informative detection of decreasing splenic function. These data suggest that spleen autotransplantation may provide a useful tool to prevent complications following splenectomy in a clinical setting.     

Cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted mice

BACKGROUND: Our department worked out a modified surgical form of spleen autotransplantation earlier, named "spleen apron method" introduced already into the clinical practice. Recently we tested the immunological changes in a group of patients autotransplanted with about 10-15% of their spleen, what was the at least always implantable amount after the severe splenic injuries. In the current work we aimed at measuring some cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted inbred mice two and eight months after the operations in order to get more unambiguous results than that we could obtain in our patients with this technique. MATERIALS AND METHODS: We divided 96 two months old Balb/c female mice into eight groups (n = 12/group). The group of controls, sham operated, splenectomized and autotransplanted animals with two and eight months of survival time after the operations. During the autotransplantation we inserted the same amount of spleen, five slices, "chips," about 10-15% of total mass of spleen, into the greater omentum similarly as it was used in the patients. The concentration of serum proteins were measured by laser nephelometry. The lymphocyte subsets were analyzed by flow cytometry. RESULTS: We found that two months after the operations the number of CD 19+ B-cells increased in the splenectomized but decreased in the autotransplanted animals. Eight months after the operations the number of both CD3+ T and CD19+ B lymphocytes decreased both in the splenectomized and autotransplanted animals compared to the controls and sham operated mice. However, the numbers of T and B cells were slightly but not significantly higher in the autotransplanted than in the splenectomized mice. The serum level of IgM was also decreased in the splenectomized and autotransplanted mice at both time points, however, eight months after the operations the concentration of IgM was significantly higher in the autotransplanted group than in the splenectomized animals. CONCLUSION: The effects of autotransplanted "chips" were different at the various ages of the animals. Additionally, they showed some immunological benefit being quantitatively in accordance to the amount of the transplanted spleen. The elevated level of serum IgM what we found in the autotransplanted mice even with this amount of transplanted spleen eight months after the operations, however, might have the potentially greatest importance compared to splenectomy. These experiments can prove that the attempts for autotransplantation may have real perspectives but their efficacy depends on the amount of the successfully transplanted (saved) mass of spleen.     

Hemorheological follow-up after splenectomy and spleen autotransplantation in mice

We previously reported on a spleen autotransplantation model in mice, with spleen function studies at 6 weeks. The present study was undertaken to investigate long-term hemorheological functions. A/J and BALB/c inbred mice were divided into four groups: control, sham surgery (SH), splenectomy (SE), and spleen autotransplantation (AU). Hematological and hemorheological parameters were determined. Leukocyte counts in the SE and AU groups were significantly higher than in controls, while hematocrit levels were markedly lower. Mean erythrocyte volume did not change significantly. Platelet counts in the AU group were significantly lower, and red blood cell deformability was significantly worse in the SE group. The AU group also had increased cell transit time, but it was less than that in the SE group. The SE group showed the highest fibrinogen levels. We conclude that there are some consistent differences in hematological parameters between splenectomy and spleen autotransplantation. These data suggest that spleen autotransplantation may partially restore hemorheological functions following splenectomy.     

The immunological studies on the splenectomy and the splenic autotransplantation in BALB/C mice

Recently, it has been demonstrated that the severe infectious diseases are often caused after splenectomy. The significance of spleen on humoral immunity has been pointed out, however the alterations of cellular immunity by splenectomy has not yet been investigated sufficiently. In this study, the effects of splenectomy on the immunological aspects were examined. And the reconstruction of immunological responses by the splenic autotransplantation was also examined. The results obtained were as follows. The antibody titers against SRBC of the splenectomized groups continued to be apparently lower than those of sham-operated groups. The IgM of the splenectomized groups showed lower values than that of the sham-operated groups, but concerning IgG, no distinct differences were noticed among these two groups. On the proliferation of peripheral lymphoid cells, the splenectomized groups from 4th week later showed a higher response than the sham operated groups. The splenic autotransplanted groups showed a similar response to the sham operated ones in the antibody production against SRBC, and the proliferations of lymphocytes. The splenic autotransplantation might be suggested to be a worthy application.     

自体脾移植联合食管下段横断术治疗肝硬化门静脉高压症的临床随机对照观察

目的　采用随机对照研究的方法观察用自体脾移植联合食管下段横断术治疗肝硬化门脉高压症的临床效果。方法　将肝功能ChildA、B级的肝硬化门脉高压症患者随机分为自体脾移植组和切脾组 ,脾移植组采用自体带蒂脾组织腹膜后移植联合改良的食管下段横断术 ,切脾组则采用脾切除联合改良的食管下段横断术 ;以患者术前的情况为对照 ,在术后 2～ 8个月观察患者的一般情况、脾扫描、肝功能、血清Tuftsin、IgM水平。结果　术后第六天切脾组死亡 1例 ,脾移植组出现再出血 1例 ;两组血清Tuftsin、IgM水平有显著性差异 (P <0 .0 5 ) ,在对肝功能的影响上无明显差异。结论　脾自体移植后能够长期存活 ,并能够维持脾脏的基本免疫功能 ,是可以在临床上推广应用的。     

Splenic tissue autotransplantation in rabbits: no restoration of host defense

BACKGROUND: The loss of spleen may increase the incidence of overwhelming sepsis. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the effectiveness of regenerated splenic tissue in preventing infection. This study explored the effectiveness of splenic tissue autotransplantation in restoring host defense. MATERIALS AND METHODS: Rabbits were divided into three groups: splenic autotransplantation, sham operation, and total splenectomy. Histomorphology, T-lymphocyte count, serum lysozyme levels, hemolysin titers, and pneumococcal clearance were observed as read-out parameters over 24 weeks. RESULTS: Histological study showed that the white pulp was poorly developed and central arterioles were missing in the regenerated splenic tissue of the autotransplanted rabbits. The weight of regenerated spleens recovered 6 months later in the splenic autotransplantation group was 11% of that in the sham operation group and was significantly less than the weight at implantation. There was no significant difference in the number of T lymphocytes or level of serum lysozyme between the three groups. A poor antibody response by the rabbits in the splenic autotransplantation and total splenectomy groups was noted after the primary intravenous administration of sheep red blood cells compared to those of sham operation group. After the challenge with type 3 pneumococci intravenously, pneumococcal clearance from the bloodstream in the splenic autotransplantation group did not differ significantly from that in the total splenectomy group, but was markedly delayed compared with that in the sham operation group. CONCLUSIONS: The low quantity and poor quality of the regenerated splenic tissue contribute to the inferior immunoprotective ability of animals autotransplanted with one-third of the original spleen. This suggests that the regenerated spleen cannot compensate for the immunological function of the original one, especially host resistance to infection.     

腹膜后自体脾移植术在肝硬变门静脉高压症治疗中的临床研究

目的　观察腹膜后自体脾移植联合食管下段横断术治疗肝硬变门静脉高压症的临床效果。方法　将20例肝功能Child A、B级的肝硬变门静脉高压症患者随机均分为自体脾移植组和切脾组。自体脾移植组采用自体带蒂脾组织腹膜后移植联合改良的食管下段横断术,切脾组则采用脾切除联合改良的食管下段横断术。以患者术前的情况为对照,在术后2~6 个月观察患者的一般情况、脾扫描、肝功能、血清促吞噬素(tuftsin)及IgM水平。结果　术后第6天切脾组死亡1例,术后第10天脾移植组出现再出血1 例。自体脾移植组术后血清tuftsin、IgM水平高于切脾组,差异有显著性意义(P<0.01),而对肝功能无明显影响。结论　腹膜后自体脾移植能维持脾脏的基本免疫功能,且能长期存活,在临床上推广应用是可行的。     

Posttraumatic autotransplantation of spleen tissue

Clinical and laboratory studies have documented high susceptibility to pneumococcal infection in asplenic humans and animals. Splenic autotransplantation has been suggested as a method of preserving function. Autotransplantation of irreparably damaged spleens in humans preserved splenic functions. Ten patients operated on for blunt abdominal trauma required unavoidable splenectomy. In each, autotransplantation of the removed spleen (roughly 50 g) was performed. Postoperative studies of splenic functions revealed disappearance of Howell-Jolly bodies from peripheral blood. Levels of IgM, which were initially significantly depressed, returned to normal and there were normal technetium Tc 99m sulfur colloid scans ten weeks after surgery. All patients are alive and healthy. Our data suggest that autotransplantation of spleen is a safe alternative method for preserving splenic function when total splenectomy is mandatory for hemostasis.     

自体脾移植联合食管横断吻合术治疗肝硬化门静脉高压症

目的 本研究比较自体脾移植联合食管横断吻合术与脾切除联合食管横断吻合术治疗门静脉高压症(PHT)的疗效.方法 将30例研究对象平均分为两组.研究组行脾次全切除加自体脾腹膜后移植及食管横断吻合术;对照组行脾切除加食管横断吻合术.于术前1周及术后6个月通过三维动态增强磁共振血管成像(3D DCE MRA)检测其门静脉主干(MPV)管腔横截面积、血流量、血流速度等门静脉系统血流动力学参数及食管胃底曲张静脉的变化,并观察自体移植脾在腹膜后的侧支血管形成与血流方向的改变.并同步检查血清肝纤维化指标及肝功能的重要指标. 结果术后两组MPV的管腔横截面积、MPV的平均流速和MPV的平均流量均比术前明显减小(P＜0.05).且术后研究组MPV的平均流速和平均流量均少于对照组(P＜0.05).两组手术后胃底曲张静脉、食管曲张静脉均消失或明显改善;两组手术前后肝功能的重要指标均差异无统计学意义(P＞0.05).研究组血清透明质酸水平在术后显著下降(P＜0.05),研究组移植脾成活良好,并建立了广泛的侧支循环. 结论自体脾移植联合食管横断吻合术是一种优于脾切除加断流术治疗肝硬化PHT的方法,且未对肝功能造成负面影响.     

自体脾移植联合食管横断吻合术治疗肝硬化门静脉高压症

目的 探讨腹膜后自体脾移植联合食管横断吻合术治疗肝硬化门静脉高压症的临床疗效.方法 将2003年1月至2006年12月收治的36例肝硬化门静脉高压症患者随机分为自体脾移植组(n=18)和脾切除组(n=18),自体脾移植组接受脾切除、食管横断吻合及自体脾移植术,脾切除组接受脾切除、食管横断吻合术.于术前及术后2～6个月定期观察两组患者的一般情况、行脾脏放射性核素扫描,同时检测肝功能、血清促吞噬素(Tuftsin)及IgM水平,并行组间及手术前后比较分析.结果 自体脾移植组患者术后2个月血清Tuftsin和IgM水平与术前比较无明显差异(P0.05),而脾切除组患者术后2个月血清Tuftsin和IgM水平较术前明显降低(P<0.05);自体脾移植术对患者肝功能无明显影响;术后2个月放射性核素扫描证实移植脾于腹膜后存活.结论 自体脾移植对保留机体脾脏免疫功能具有重要价值,腹膜后自体脾移植联合食管横断吻合术治疗肝硬化门静脉高压症的临床效果确切,值得推广应用.     

Phagocyte function after splenic autotransplantation

This study was designed to examine the role of splenectomy and autotransplantation with regard to the leukocyte/differential cell counts and the function of peripheral blood phagocytes. Eleven groups of 40 Wistar male rats in each group either underwent total splenectomies or sham operations. The splenectomized groups underwent autotransplantations with 10% through 90% of the weight of the intact spleen. The leukocyte count and the oxidative burst response of the blood leukocytes were measured in each group. It was shown that a total splenectomy did not alter the leukocyte/differential cell counts. Furthermore, the blood picture remained basically unchanged after an autotransplantation with 10% through 90% of the weight of the intact spleen. The phagocyte oxidative burst response was measured by chemiluminescence. The chemiluminescence response of these cells was reduced after a total splenectomy. The phagocyte oxidative burst response returned to normal levels following an autotransplantation. There was no correlation between the amount of autotransplanted spleen and the degree of the oxidative burst response. These findings indicated that a splenectomy results in a diminished phagocyte oxidative burst response and that a spleen autotransplantation returns this function to normal levels.     

进展期近端胃癌根治术联合脾脏切除术适应证的选择

目的探讨进展期近端胃癌根治术联合脾脏切除术的合理适应证。方法选取我院自2005年3月至2008年2月期间50例进展期近端胃癌行全胃切除患者为研究对象,根据脾脏是否切除分为切脾组(n=18)和保脾组(n=32),比较2组患者间手术时间、住院时间、术后并发症以及脾门淋巴结转移情况。结果切脾组手术时间、住院时间和膈下感染发生率均明显高于保脾组(P0.05)。切脾组与保脾组术后病理检查出的第10、11组淋巴结转移率的差异无统计学意义(P0.05)。结论进展期近端胃癌根治术时,脾脏和脾血管受侵是联合脾脏切除的合理适应证。     

Relative merits of partial splenectomy, splenic reimplantation, and immunization in preventing postsplenectomy infection.

Partial splenectomy, splenic autotransplantation, and immunization with pneumococcal vaccine have been reported to protect patients against overwhelming postsplenectomy infection, and this study was undertaken to evaluate these therapeutic alternatives. For this purpose 136 rats were divided into experimental groups: 34 controls, 34 splenectomy, 34 partial splenectomy, and 34 splenic autotransplantation animals. Five weeks after operation, two-thirds of the animals were immunized with killed pneumococci. The effects of operation and immunization were studied by challenging the animals intravenously with pneumococci. Pneumococcal antibody titers were determined, and phagocytic uptake of pneumococci by the spleen and liver was measured. Immunization impressively increased the survival rate in all groups. At low-challenge doses autotransplantation prolonged survival. At higher-challenge doses only partial splenectomy increased survival. Partial splenectomy and control animals had higher antibody titers than did splenectomy and autotransplantation rats. Animals with the highest antibody titers had the greatest splenic and hepatic phagocytic uptake of pneumococci. Partial splenectomy was more efficient in removing pneumococci than was autotransplantation. Thus immunization is one of the most important factors contributing to survival after splenectomy. Partial splenectomy is preferable to splenic autotransplantation because it is associated with higher antibody titers after immunization, better pneumococcal splenic uptake, and improved survival rates.     

Comparative investigation of the hematologic and immunologic effects of splenectomy and spleen autotransplantation in mice

We have analysed the hematological and immunological effects of the Furka's spleen chip method of autotransplantation in Balb/c mice two and eight months after splenectomy and autotransplantation. Our results showed that the autotransplants had positive effect on the partial restoration of the damaged elements of both cellular and humoral immunity appearing after splenectomy. However, these beneficial effects may have some dependence of time and species.     

The value of partial splenic autotransplantation in patients with portal hypertension: a prospective randomized study.

HYPOTHESIS: Splenic autotransplantation plays a role in preserving immune function of the spleen in patients with portal hypertension and liver cirrhosis. DESIGN: Prospective randomized study. SETTING: University hospital. PATIENTS: Twenty patients (19 men and 1 woman; aged 33-80 years) suffering from portal hypertension and liver cirrhosis were randomly allocated into 2 groups. Each group consisted of 10 patients. INTERVENTIONS: All patients underwent modified Sugiura operation. In the control group, splenectomy was performed, while partial splenic autotransplantation into the retroperitoneal space was additionally completed in the splenic autotransplantation group. MAIN OUTCOME MEASURES: Serum tuftsin and IgM were measured preoperatively and 2 months after surgery. Dynamic scintigraphy with technetium Tc 99m-labeled heat-damaged erythrocytes was performed at 2-month intervals during the 8-month follow-up. RESULTS: There was no statistical difference in the mortality of the groups. The preoperative levels of serum tuftsin and IgM showed no statistical difference between groups. However, although these measures had decreased remarkably in the control group 2 months after operation (P<.001 for serum tuftsin; P =.04 for serum IgM), they remained stable in the splenic autotransplantation group (P =.25 for serum tuftsin; P =.12 for serum IgM). Four patients within the splenic autotransplantation group showed positive scanning of the transplanted splenic fragment during follow-up, whereas there was no positive scanning in the control group. CONCLUSION: Our results suggest that partial splenic autotransplantation can preserve immune function of the spleen, as measured by serum levels of tuftsin and IgM, in patients with portal hypertension and liver cirrhosis.     

The Effects of Splenectomy and Splenic Autotransplantation on Plasma Lipid Levels

Purpose: Atherosclerosis observations after splenectomy for trauma and hypersplenism suggests a possible role for the spleen in lipid metabolism. The authors examined the effects of splenectomy on serum lipids in rats and also cholesterol-fed rats with experimental atherosclerosis. Methods: This study was designed on rats. The rats were divided into five groups: splenectomy, normal diet (SP-N, n: 8), splenectomy, cholesterol-fed groups (SP-C, n: 8), splenic autotransplantation after splenectomy, normal diet (SA-N, n: 8), splenic autotransplantation after splenectomy, cholesterol-fed groups (SA-C, n: 8) and sham groups (n: 8). Total triglyceride, total cholesterol, HDL (high-density lipoprotein), LDL (low-density lipoprotein), and VLDL (very low-density lipoprotein) levels were determined in 40 rats. The rats were classified into five groups based on the surgical procedures. The spleens were removed and then the rats were fed a normal diet in Group SP-N (n = 8). The spleens were removed and then the rats were fed a diet containing 1% cholesterol in Group SP-C (n = 8). Splenectomy and splenic autotransplantations were performed and then the rats were fed a normal diet in Group SA-N (n = 8). Splenectomy and splenic autotransplantations were performed and then the rats were fed a diet containing 1% cholesterol in Group SA-C (n = 8). The rats were sham-operated in the control group (Group S, n = 8). An active splenic function was shown in rats that underwent splenic autotransplantation in both groups by using Technicium 99 m sulphurcolloide sintiscan on day 30. Blood lipid levels were repeated 6 months later. Results: There was no difference between pre- and postoperative lipid levels in the sham group and SA-N group (p >.05). All lipid levels including HDL were increased significantly in SP-C group (p <.05). Also VLDL and total tryglyceride levels were increased significantly in SP-N and SA-C groups (p <.05). Conclusions: This study showed that the spleen might have an important effect on lipid metabolism and splenic autotransplantation may be protective in conditions with increased lipid levels.     

Optimal site and amount of splenic tissue for autotransplantation.

Clinical and basic studies have documented a high susceptibility to pneumococcal infection in asplenic humans and animals. It has been suggested that autotransplantation of splenic tissue might be a method of providing host resistance when total splenectomy is necessary. However, the effect of splenic autograft has remained controversial. This study was performed to evaluate the most effective site and amount of splenic autograft using rats. Rats were divided into five groups for the purpose of determining the site of splenic autotransplantation: splenectomy, sham operation, implantation into the omental pouch, intraperitoneal implantation, and intramuscular implantation. For determining the amount for autotransplantation, the rats were divided into seven groups: splenectomy, sham operation, and implantations of 25, 50, 100, 200, or 300 mg of splenic tissue. All animals were challenged with Streptococcus pneumoniae type 6, 16 weeks after surgery. Howell-Jolly bodies appeared postsplenectomy, but disappeared in the implanted rats 16 weeks after the operation. Histologically, the implanted tissue was indistinguishable from that of a normal spleen. Pneumococcal clearance from the bloodstream and survival rate were significantly higher in rats implanted in the omental pouch as compared with splenectomized rats. Intraperitoneal and intramuscular implanted rats did not show a significant difference from the splenectomized rats. More than 50% of splenic tissue for autograft showed a significant increase in pneumococcal clearance and survival rate as compared with that of splenectomized rats. It was suggested that the most effective site of autotransplantation is the omental pouch and approximately 50% of the whole spleen would be necessary for prevention from sepsis.     

Distribution of peripheral blood cells in mice after splenectomy or autotransplantation

Our aim was to compare the distribution changes of peripheral leukocytes and erythrocytes in splenectomized and spleen-autotransplanted BALB/c female mice (n = 96), 2 and 8 months after surgery. In total, there were eight groups of animals: splenectomy, autotransplantation, sham, and untreated controls at both time points. We used the spleen-apron method of Furka et al. (Khirurgiia (Mosk) 1989;9:125-127), inserting five spleen chips into the greater omentum, for autotransplantation. Quantitative and qualitative blood cell counts and the phagocytic activity of cells (stimulated with zymosan) were determined. In splenectomized animals, the number of neutrophils significantly increased 8 months after surgery. The greatest phagocytic activity of neutrophils, however, was observed in autotransplanted animals of the same age. In splenectomized animals, erythrocyte volumes were significantly higher in the second postoperative month, but normalized by the eighth month. In conclusion, spleen autotransplantation has some beneficial effects, including clearing erythrocytes and preserving the phagocytic activity of neutrophils in peripheral blood.     

脾脏切除对门脉高压症病人NK细胞活性的影响

门脉高压症瘀血肿大的脾脏是否具有免疫功能尚存有分歧。本文通过对保留脾脏的远端脾肾静脉分流手术病人和切除脾脏的门奇静脉断流手术病人的外周血NK细胞活性的测定,对照分析了脾切除对门脉高压症病人NK细胞活性的影响。结果表明:门脉高压症病人NK细胞活性显著低于正常人;脾切除后NK细胞活性呈渐进性升高,而保留脾脏的病人NK细胞活性无明显变化,术后两周脾切组显著高于保脾组(P<0.01)。提示门脉高压症病人瘀血肿大的脾脏对机体非特异性免疫功能有抑制作用,睥切除术后早期可改善门脉高压症病人的NK细胞活性。其远期效果有待随访。     

脾大部切除及食管横断吻合术治疗肝硬化门静脉高压症的血流动力学研究

目的 评价脾大部切除、残脾腹膜后包埋及食管横断吻合术对肝硬化门静脉高压症患者门静脉血流动力学的影响.方法 将40例门静脉高压症患者随机分为研究组和对照组,每组20例.均进行食管横断吻合术,对照组做全脾切除,研究组保留部分带血管蒂脾脏移植于腹膜后.手术前后用三维动态增强磁共振血管成像测量门静脉主干的管腔横截面面积、血流量、血流速度和流向;观察自体移植脾在腹膜后的血供及侧支循环.结果 两组术前均存在胃底食管曲张静脉,术后6个月MRA复查均消失或改善.术后6个月两组门静脉主干的管腔横截面积明显减少[研究组(1.81±0.73)cm~2比(1.20±0.52)cm~2,P＜0.01;对照组(1.78±0.52)cm~2比(1.30±0.12)cm~2,P＜0.01];术后两组门静脉主干的平均流速均下降[研究组(9.86±0.10)cm/s比(7.06±1.92)cm/s,P＜0.01;对照组(10.0±0.6)cm/s比(8.2±2.4)cm/s,P＜0.01],且研究组少于对照组(P＜0.01);术后两组门静脉主干的平均流量均下降[研究组(15.0±1.9)ml/s比(10.5±2.7)ml/s,P＜0.01;对照组(14.9±2.1)mI/s比(11.6±2.1)ml/s,P＜0.01],且研究组少于对照组(P＜0.05).移植脾在腹膜后成活,并建立了广泛的侧支循环.结论 脾大部切除、带血管蒂残脾腹膜后移植及食管横断吻合术治疗肝硬化门静脉高压症不仅保留了脾脏的功能,而且具有断流和分流为一体的联合性术式的作用.