Adrenal function in different phases of the estrous cycle in the domesticated silver fox, Vulpes fulvus

Experiments were made on silver foxes from a population which had been selected for 10 to 15 generations for the domestic behaviour and on animals from a control, unselected population. In females from both populations, studies were made of the level of 11-OHCS in the blood serum, in vitro production of 11-OHCS, the size of fascicular zone in the adrenal cortex, the volume of cellular nuclei and nucleoli, as well as the reaction of the adrenals in vitro to 2 doses of ACTH (1 and 5 units/g of the adrenals) during anestrus, proestrus and estrus. In control females, all the characters investigated significantly increased from anestrus to proestrus. In domesticated females, no changes in the production of 11-OHCS in vitro or changes in morphological features of the fascicular zone were observed in the course of estrous cycle. During proestrus, the adrenals of the domesticated animals were not able to increase the production of 11-OHCS in vitro after application of ACTH. The decrease in the reactivity of the adrenals to the effect of ACTH is presumably the main cause why in unselected females the adrenal are not activated during proestrus. Therefore, in the course of selection for the domestication type of behaviour, species specific dynamics of activation of the adrenals during estrous cycle is lost.     

Effect of an oral contraceptive on the lysine-8-vasopressin test in the diagnostic evaluation of pituitary function.

A comparative study was performed on the plasma 11-hydroxycorticosteroids (11-OHCS) responses to lysine-8-vasopressin (LVP) before and during ingestion of an oral contraceptive wieh oestrogenic activity. A total of 19 healthy women with a normal menstrual cycle and normal plasma 11-OHCS content in blood samples at 8 a.m. and 4 p.m. were included in the study. The results show that the mean response of plasma 11-OHCS to the administration of LVP was equal in magnitude before and during ingestion of the contraceptive, although the baseline value of 11-OHCS doubled as a result of the treatment. On the other hand, during contraceptive treatment the increase of plasma 11-OHCS concentration after the administration of LVP was somewhat slower, reaching its peak later than before treatment. The noticeable variation in individual responses of plasma 11-OHCS to LVP increased during the treatment. Three subjects showed weak responses to LVP, although the adrenocortical responses to exogenous ACTH were quite normal. The inidvidual responses to LVP before and during ingestion of an oral contraceptive with oestrogenic activity are discussed.     

Function of the adrenal cortex during elaboration of the passive avoidance conditioned reflex in immature rats

Plasma 11-hydroxycorticosteroid (11-OHCS) levels were measured in 30-day-old rats by fluorometry during passive avoidance (PA) learning by means of a single electric footshock. In contrast to the data obtained in adult animals, pre-exposure of young rats for 7 days to the experimental environment (over 3 min daily) resulted in elevation of the basal 11-OHCS levels and in the lack of distinct changes in the hormonal background after placing the young rats into a chamber. As in previous experiments on adult rats, one day after PA learning the 11-OHCS levels were significantly lower in young rats displaying PA than in the animals which did not exhibit PA behavior. Five days after PA training these differences in adrenocortical reactivity disappeared, as was the case in adult animals.     

Changes of maternal and fetal pituitary-adrenocortical functions during human labour

The results of maternal 11-hydroxycorticoid (11-OHCS) determination on blood obtained from four stages of 44 spontaneous labour cases showed a strong and time-related maternal adrenocortical activation during vaginal delivery. The high 11-OHCS level in the cord blood and the increase of the 11-OHCS concentration in the amniotic fluid suggested the same response of the fetal adrenal. However, in 14 cases of elective caesarean sections the 11-OHCS levels in the cord blood and the amniotic fluid did not rise in spite of the elevated maternal plasma 11-OHCS content.     

Effect of Resection of Lung Tumours on the Steroid Abnormalities in Patients with Lung Cancer

The urinary excretion of androsterone, aetiocholanolone, total 17-oxosteroids, and 17-hydroxycorticosteroids (17-OHCS) was measured in 40 patients with lung cancer three days before resection and again 10-15 days after resection of their lung tumours. There was a significant postoperative increase in the excretion of 17-OHCS but a significant decrease in the excretion of androsterone and aetiocholanolone, resulting in an increase of the preoperative abnormalities in steroid excretion in these patients. Since there was no change in steroid excretion towards normal after resection of the lung tumours, it seems that the steroid abnormalities found in lung cancer are not the effect of the presence of the lung tumours. As the excretions of 17-OHCS and 11-deoxy-17-oxosteroids change in opposite directions after resection, it is suggested that a dissociation of factors that control the excretion of these two groups of steroids takes place as a response to surgical stress in patients with lung cancer.     

Passive immunization of rams (Ovis aries) against GnRH: effects on antibody titer,serum concentrations of testosterone,and sexual behavior

The effect of immunoneutralization of gonadotropin-releasing hormone (GnRH) on serum concentrations of testosterone and sexual behavior was evaluated in sexually mature male sheep. In Experiment 1, GnRH1 rams (n=16) were passively immunized against GnRH (300 ml antiserum), control rams were either passively immunized against keyhole limpet hemocyanin (KLH, n=15) or surgically castrated (Wethers1, n=4). Sexual performance of the rams was assessed weekly for 3 weeks before and 6 weeks after immunization, using ovarihystertomized ewes actively immunized against GnRH. Experiment 2 evaluated the effects of repeated immunization. Rams were immunized with two aliquots (400 and 300 ml, respectively) of anti-GnRH sera (GnRH, n=5) or normal sheep serum (NSS, n=4), 2 weeks apart. Surgically castrated animals were used as a second control group (Wethers2). Administration of anti-GnRH sera, but neither anti-KLH nor NSS sera, resulted in marked reduction (P<0.05) in serum concentrations of testosterone. Sexual behavior was not consistently affected by administration of one aliquot of anti-GnRH sera, however repeated immunizations resulted in more persistent reduction in serum concentrations of testosterone and more consistent suppression of sexual behavior.     

Induction of a cellular immune response to a foreign antigen by a recombinant Listeria monocytogenes vaccine.

A recombinant strain of Listeria monocytogenes that stably and constitutively expresses Escherichia coli beta-galactosidase was used as a live vaccine vector. BALB/c mice were immunized orally or parenterally with the recombinant L. monocytogenes, and their cellular and humoral immune responses to beta-galactosidase were measured. Spleen cells taken 1 week after oral inoculation or 5 weeks after oral or parenteral inoculation (with a boost at 4 weeks) showed beta-galactosidase-specific CTL responses. The CTL line derived from mice immunized i.p. was also shown to be class I restricted and Thy-1.2+, CD8+, and TCR alpha beta+. All mice immunized with the recombinant L. monocytogenes had positive delayed-type hypersensitivity responses to heat-killed L. monocytogenes, but only 15% had a positive delayed-type hypersensitivity reaction to beta-galactosidase. Individual serum samples from mice immunized i.p. or i.v. were tested for antibody to beta-galactosidase. Approximately 11% had low positive titers for beta-galactosidase antibodies. These results demonstrate that both oral and parenteral immunization with recombinant L. monocytogenes results in a cellular immune response to the foreign protein, which is primarily a specific CD8+ CTL response.     

Effect of active immunization against androstenedione on ovarian function in dairy cows and prepubertal gilts

Two experiments were performed to assess the effects of immunization against androstenedione on ovarian function in cows during the first 100 days after calving and in gilts at the onset of puberty.Seven Holstein-Friesian cows were immunized against androstenedione-11α-human serum albumin (AND-HSA) at 3 weeks before and at 3 weekly intervals after calving and compared with ten cows immunized against HSA. Titres against androstenedione were significant but low and variable (1: 280-1: 4). Two cows with the highest titres exhibited abnormal ovarian function which could be attributable to treatment. One cow developed a large follicular cyst and nymphomania. The other cow exhibited mild super ovulation in her first cycle after calving and multiple ovulations during later cycles. Other cows in the AND-HSA group also had a tendency to develop larger than normal follicles and follicular cysts.Groups of twelve prepubertal gilts were immunized against either androstenedione 11α-hemisuccinyl bovine serum albumin (AND-BSA) or BSA at 100, 121 and 142 days of age. Titres against AND were significant but were low and variable (1: 2500-1: 10). There was no effect of immunization against AND on onset of ovulation, puberty or ovulation rate in the first or second ovulatory cycles.     

Progesterone secretion by adrenal glands of hamsters and comparison of ACTH influence in rats and hamsters.

Studies were designed to determine a) if adrenal glands of hamsters secrete progesterone (PROG), b) the effects of adrenocritocotropin (ACTH) administration on adrenocortial function of rats and hamsters under the surgical conditions necessary for collection of adrenal venous blood from the left renal vein, and c) the effects of blood loss during sample collection. PROG was quantitated by the competitive protein-binding method after extraction and separation by sephadex LH-20 column chromatography. The presence of interfering quantities of androstenedione necessitated two column chromatographic steps. Glucocorticoids (11-OHCS) were determined fluorometrically. PROG was detected in adrenal venous plasma of female hamsters. The PROG concentration and secretory rate were 91 +/- 12 ng/ml and 4 +/- 1 ng/min, respectively, while the peripheral plasma level of the same animals was 2 +/- 0.2 ng/ml, indicating that the adrenal glands of female hamsters are capable of secreting PROG. ACTH administration increased PROG secretory rates in both hamsters (3 +/- 1 to 14 +/- 3 ng/min) and rats (62 +/- 9 to 152 +/- 32 ng/min) on estrus, as well as increasing the 11-OHCS secretory rate of hamsters (16 +/- 1 to 33 +/- 4 ng/min), but not of rats. The greater increase in PRCC than in 11-OHCS secretion may be related to excess PROG formation relative to the capacity of the 17alpha- or 21-hydroxylating enzyme systems. The adrenal venous PROG concentration and secretory rate of female hamsters infused with 10% dextran while collecting adrenal venous blood did not differ significantly from those of the non-infused animals, suggesting that this amount of blood loss (1 ml) does not influence PROG secretion.     

Potentials of Shigella flexneri Y strain TSF21 as a candidate vaccine against shigellosis: safety, immunogenicity and protective efficacy in Bonnet monkeys

A thymine-requiring and temperature-sensitive mutant of Shigella flexneri Y was tested in Bonnet monkeys for safety, immunogenicity and protective efficacy. A dose of 10(11) cells when fed orally mimicked natural infection in having invaded epithelial cells, but was otherwise clinically non-reactogenic. Animals immunized with two oral doses, each dose consisting of 1 x 10(11) mutant bacteria, were fully protected when challenged, with respect to the lack of any clinical symptoms or detectable histological abnormalities in the intestinal mucosa. Unimmunized animals when similarly challenged developed frank dysentery and the intestinal mucosa showed severe histological abnormalities. Titres of serum antibodies increased by about 11-fold of the base level in animals immunized with a dose of 10(11) cells, but not with lower doses. The challenge bacteria appeared to be phagocytised by macrophages. In some monkeys of a particular group, congestive patches were seen in the stomach, but not in any other part of the gut, after the animals were fed with the virulent parent strain. The lesions were relatively severe in the immunized groups of animals.     

Resistance to Naegleria fowleri infection passively acquired from immunized splenocyte, serum or milk

A pathogenic free-living amoeba, Naegleria fowleri, causes primary amoebic meningoencephalitis to human and experimental animals. This infection is rare, but the mortality is very high. Nowadays, drug treatment or active immunization of human or mice are being tried with partial effectiveness. This study shows passive immunization effect by transfer of immunized spleen cells, serum, or milk from immunized mother in mouse experimental model. Young BALB/c mice were immunized intraperitoneally with 2-3 X 10(6) trophozoites of N. fowleri, and spleen cells and sera were collected for injection to recipient mice. There were seven transfer groups, i.e., immunized mouse serum, spleen cells, serum and spleen cells, normal mouse serum, spleen cells, serum and spleen cells, and control group. Three days later, BALB/c mice were inoculated with 1 x 10(4) trophozoites of N. fowleri intranasally. After infection, decreased mortality and prolonged survival time of mice were noted in immunized groups compared with non-immunized control group. The groups injected with immunized spleen cells or normal serum showed lower mortality than that of controls but showed no changes of serum IgG level. The groups injected with immunized serum or normal spleen cells showed increased serum IgG level after immunization but hundred percent mortality was observed. Mother mice were immunized intraperitoneally with 2-3 X 10(6) trophozoites of N. fowleri at the end of pregnancy and weaning period. Soon after the delivery, litters born of non-immunized mother were matched with immunized mother for feeding immune milk. After three weeks, the litters were infected with 1 X 10(4) trophozoites of N. fowleri or sacrificed for serum collection to measure the IgG levels.(ABSTRACT TRUNCATED AT 250 WORDS)     

SARS-CoV-2重组S1和S蛋白疫苗诱导保护性免疫的研究*

目的:评价新型冠状病毒(SARS-CoV-2)重组S1蛋白和S蛋白疫苗对SARS-CoV-2的免疫保护效果。方法:将SARS-CoV-2重组S1蛋白和S蛋白分别联合氢氧化铝佐剂以0.1 μg/只、1 μg/只、5 μg/只、10 μg/只不同剂量接种6~8周BALB/c纯系健康雌性小鼠。第二次免疫后采血通过酶联免疫吸附试验(ELISA)检测血清中IgG抗体效价,通过假病毒中和试验比较免疫小鼠血清对SARS-CoV-2野生型株(WT)、英国株(B.1.1.7)、巴西株(P.1)、印度株(B.1.617.2)、Mu毒株(B.1.621)和南非株(501Y.V2-1)六种假病毒毒株中和活性效价,取脾细胞通过酶联免疫斑点技术(ELISpot)检测免疫小鼠的细胞免疫水平。结果:SARS-CoV-2重组S和S1蛋白都能诱导小鼠产生较强的IgG抗体水平。免疫S1蛋白的小鼠血清对SARS-CoV-2野生型株、英国株、巴西株有明显的中和活性,免疫S蛋白的小鼠血清除了对SARS-CoV-2野生型株、英国株、巴西株有明显中和活性之外,对印度株也有明显的中和活性,两种蛋白质免疫的小鼠血清均对野生型株中和效果最强。S蛋白免疫的小鼠脾细胞能够显著诱导出γ干扰素(IFN-γ)和白介素-4(IL-4)的产生。S蛋白诱导产生的IgG抗体、中和抗体、细胞免疫水平均高于S1。结论:SARS-CoV-2重组S蛋白疫苗能够诱导产生较强的保护性免疫应答。     

Resistance of mice with active and maternally passive immunity against Sendai virus

Resistance of mice with active and maternally passive immunity to Sendai virus infection was investigated. Mice with active immunization were convalescent ones from intranasal infection with 10(3) TCID50 of the virus [CT], ones immunized with 4 weekly intranasal injections of about 4 X 10(3) hemagglutinating units (HAU) of formalin-inactivated virus (FV) [IN], or ones immunized with 4 weekly intraperitoneal injections of about 2 X 10(3) HAU of FV [IP]. The serum neutralizing (NT) antibody titers ranged 1 : 10 to 1 : 20 in CT and IN, and 1 : 20 to 1 : 40 in IP at 4 weeks after initial immunization. NT antibody in the lung lavage was detected only in IP in 3/5. After intranasal challenge infection with 10(6) TCID50 of the virus, little or no gross lung lesion, no virus recovery and no body weight loss were observed throughout experiment, in these 3 immunized groups, whereas, control mice showed lung lesions in 4/5 (7 days), virus recovery in 4/5 at 3 days and in 1/5 at 7 days post-challenge, and body weight loss. In additional histological study, bronchiolar epithelial methaplasia and alveolar septal thickening, which were characteristic findings in non-immunized infected mice, were not observed in mice immunized with 2 biweekly injection of about 250 HA of FV and then challenged. Maternal immunity was investigated in offsprings from convalescent dams infected with 10(3) TCID50 at mating [mCT] and from dams immunized intraperitoneally with 4 X 10(3) HAU of FV at 0, 1 and 2 weeks after mating and, 1 and 2 weeks after parturition [mIP]. Serum NT antibody was not detected in mCT, but the titers ranging 1 : 20 to 1 : 40 were detected in mIP. After intranasal challenge of mCT, mIP and offsprings from non-immune mice with 10(3) TCID50 of the virus, virus recovery on day 3 was 2/4, 1/4 and 3/3, and incidence of total lung lesions on day 7 and 12 was 11/21, 2/13 and 16/16, respectively. Body weight gain was suppressed slightly in the immune groups but markedly in the non-immune control.     

The plasmatic 11-hydroxy-corticosteroid response in the human to a synthetic octadecapeptide corticotrophin (C-41795-Ba).

The adrenal response as measured by the plasmatic changes in 11-hydroxy-corticosteroid (11-OHCS) concentration following the administration of a synthetic octadecapeptide (C-41795-Ba) was compared in 17 normal young males with that obtained with the commercially available tetracosactrin. C-41795-Ba polypeptide shows a higher potency than tetracosactrin. Its effects are longer lasting when administered intravenously which indicates a significant local inactivation of the polypeptide.     

The excretion of 11-hydroxycorticosteroids by rats during exposure to altitude

The effect of exposure to 5,500 m and 7,300 m simulated altitude on rat adrenal glands was assessed by the measurement of urinary 11-hydroxycorticosteroids (11-OHCS). Since rat urine contains large amounts of non-corticosteroid fluorogens, a modification was introduced into the conventional fluorimetric method. At 5,500 m altitude increased excretion of 11-OHCS was confined to the first 24 hr. At 7,300 m the excretion of 11-OHCS continued at elevated levels throughout the exposure period. The excretion of catecholamines was also increased in rats during exposure to altitude. Since changes in the levels of excreted adrenaline and body weight were comparable with changes in 11-OHCS excretion at both altitudes, in the rat urinary 11-OHCS levels appear to constitute an index of the stress imposed by high altitudes.

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Zusammenfassung Die Wirkung von 5.500 m und 7.300 m simulierter Höhe auf die Nebennieren der Ratte wurde überprüft durch Messung der 11-Hydroxykortikosteroide (11-OHCS) im Harn. Da Rattenharn grosse Mengen von Fluorogenen enthält, die keine Kortikosteroide sind, wurde die konventionelle fluorimetrische Methode modifiziert. In 5.500 m Höhe bestand eine erhöhte 11-OHCS Ausscheidung nur in den ersten 24 Stunden. In 7.300 m Höhe wurde eine erhöhte Ausscheidung von 11-OHCS während der gesamten Expositionsperiode von 14 Tagen beobachtet. Die Ausscheidung der Katecholamine war während der Periode in den beiden Höhen ebenfalls erhöht. Da die Veränderungen der Adrenalinausscheidung und des Körpergewichts vergleichbar waren mit den Veränderungen der 11-OHCS Ausscheidung scheint der 11-OHCS Gehalt im Harn als Index für das Ausmass der Höhenbelastung bei Ratten geeignet zu sein.

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Resume On a examiné l'effet de l'exposition à des altitudes simulées de 5,500 m et de 7,300 m sur les surrénales des rats en mesurant le taux de 11-hydroxycorticostéroïde (11-OHCS) dans l'urine. Comme cette dernière contient de grandes quantités de substances fluorogènes qui ne sont pas des corticostéroïdes, on a modifié la méthode conventionelle de mesure fluorimétrique. A 5.500 m d'altitude, on ne constate une sécrétion accrue de 11-OHCS que durant les premières 24 heures d'exposition. A 7.300 m d'altitude par contre, la sécrétion accrue a duré jusqu'à la fin de l'expérience, c'est-à-dire 14 jours. On a constaté en outre une sécrétion accrue de catécholamine dans les deux cas et durant toute la période. Comme les variations de poids et de sécrétion d'adrénaline étaient en relation directe avec celles de 11-OHCS, le taux de ce dernier dans l'urine semble être un bon indice pour déterminer la contrainte d'altitude chez les rats.

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DCIEM Research Paper 821.     

Clinical chronopharmacology of ACTH 1-17. I. Effects on plasma cortisol and urinary 17-hydroxycorticosteroids

The aim of the investigation was to study the effects of ACTH 1-17 on both plasma cortisol and urinary 17-OHCS in health adult young males with regard to the time (clock hours) at which this polypeptide was injected. Eight healthy adults (males from 18-30 years) volunteered for the study. They were synchronized with a diurnal activity from 0700 to 0000 and a nocturnal rest. Each week, during 6 consecutive weeks (January 19 to February 25, 1980), a 3-day test was performed on Saturday, Sunday and Monday. On Sundays 3 control-tests and 3 ACTH-tests were programmed during which either saline or 100 micrograms ACTH 1-17 were injected i.m. at respectively 0700, 1400 and 2100. During each 3 day-test (72 h) the urinary excretion of 17-OHCS was determined every 4 h at fixed clock hours. In addition, on Sundays, venous blood was sampled prior to control or ACTH injections at respectively 0700, 1400, and 2100 and 20, 40, 60, 90, 120, 150 and 180 min thereafter. Plasma cortisol (radioimmunoassay) was determined in samples thus collected. Both conventional and cosinor methods were used for statistical analyses. A strong and statistically significant rise of plasma cortisol was observed after all of the ACTH 1-17 injections. The obtained mean response curves were observed after all of the ACTH 1-17 injections. The obtained mean response curves were similar in form and parallel. The highest plasma cortisol curve corresponded to ACTH injected at 0700, the lowest to ACTH injected at 2100. The curve corresponding to ACTH injected at 1400 went in-between. The 24-h urinary excretion of 17-OHCS after ACTH 1-17 was approximately 4 times greater than the control value when injected at 0700, approximately 3 times greater than control when injected at 1400 and only twice greater than control when injected at 2100. In terms of changes in plasma cortisol and 17-OHCS the greatest best benefit of ACTH 1-17 is achieved when this polypeptide is injected at 0700, rather than at 1400 or 2100 in diurnally active subjects.     

人巨细胞病毒磷蛋白65及糖蛋白B在大肠杆菌中的高效表达、免疫原性研究及初步应用

实验成功的构建了含有pp65与gB主要抗原决定簇基因的表达质粒pET-pp65、pET-gB;表达产物通过SDS-聚丙烯酰胺凝胶电泳、免疫印迹、间接酶联免疫等一系列鉴定试验进行了分析。表达蛋白经初步纯化后免疫BALB/c小鼠,经0、2、4周免疫三次,免疫后2、4、6周眼眶采血收集抗血清,进行中和试验和ELISA检测。动物试验表明两种表达蛋白具有免疫原性,并可在小鼠体内诱导产生特异性抗体,其中用重组gB蛋白免疫小鼠后得到的抗血清在体外试验中能抑制天然病毒感染细胞。同时将这两种表达蛋白作为诊断用抗原,对临床94份血清进行检测,证实pp65具有较好的特异性,为今后研制HCMV诊断试剂提供了科学依据。     

Maintenance of plasma-derived proteins at much lower concentrations in the uterine lumen of the rabbit: a kinetic study of passage.

Human serum albumin (HSA) and human gamma globulin (HGG) in serum and uterine fluid of nonpregnant rabbits at various times after an i.v. injection (100 mg/kg) were measured by a radial immunodiffusion test using specific antisera. The HSA concentration in uterine fluid rose to a peak at 12 hr when it was 11% of the serum concentration and then declined, whereas HGG reached a peak at 18 hr (3.2% of serum level) and decreased thereafter. The HSA passed 2 1/2 times faster than HGG, but both proteins equilibrated with uterine fluid in about 12-18 hr. Steady state levels of HSA and HGG indicated that uterine fluid: serum ratios were 1:10 and 1:20, respectively. Similar ratios were found for total protein and rabbit serum albumin (1:10) and rabbit gamma globulin (1:20). Therefore, except when there is a local immune response, the uterine lumen contains only about 5% of the serum antibody concentration. Available data in the mouse, rat and dog also indicate disparity between serum and uterine fluid protein levels.     

Plasma 11-Hydroxycorticosteroid and Growth Hormone Levels in Acute Medical Illnesses

Adrenal cortical response in acute medical illness has been studied by measuring the plasma 11-hydroxycorticosteroid (11-OHCS) concentration in 178 patients. Those with unbalanced diabetes, acute infections, and severe myocardial infarction had high levels. The results obtained suggest that in a patient with a severe infection and hypotension a plasma 11-OHCS level of less than 15 μg./100 ml. indicates an inadequate adrenal cortical response, and one patient with septicaemia and temporary adrenal cortical insufficiency is described. Growth hormone levels were increased in patients with severe diabetic ketosis but not in those with hyperosmolar non-ketotic diabetic coma.     

猪胸膜肺炎放线杆菌血清1型RTX毒素I的N-端表达多肽具有良好的免疫原性

ApxI外毒素是猪胸膜肺炎放线杆菌(APP)最重要的毒力因子,为了研究其N端多肽的免疫原性,分别将apxIA基因的全长编码区(apxIA,3146bp)及其5′端1140bp的片段(apxIA5)克隆到原核表达载体pET28a,经IPTG诱导后在大肠杆菌中实现了表达,表达产物ApxIA和ApxIAN均以包涵体的形式存在,Westernblot检测证实两种表达产物均具有免疫反应性。将纯化的重组蛋白(rApxIA和rApxIAN)和提取的天然毒素ApxI(nApxI)分别经腹腔免疫BALBc小鼠,于免疫前、免疫2周和4周后分别检测了ELISA抗体和毒素中和抗体水平,结果表明,rApxIAN免疫组的ELISA抗体显著低于rApxIA免疫组和nApxI免疫组,但rApxIAN免疫组血清中和试验中测定的溶血素单位与rApxIA及天然nApxI免疫组没有显著差异。第二次免疫2周后,用1个LD50的APP血清1型J101株和2型标准菌株攻击试验动物,rApxIAN免疫组对血清1型和2型菌株的保护率分别为80%和100%。