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@#Intractable hemorrhagic cystitis is an uncommon but significant treatment complication of concurrent chemoradiation therapy for cervical cancer. Alum instillation is regarded as a safe and effective option for its treatment. This case presentation will discuss a patient who presented with postradiation cystitis and was treated with alum irrigation. The aim of this report is to offer alum irrigation as a management option for intractable hematuria.
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Cancer SurvivorsABSTRACT
Resumen ANTECEDENTES: Las quemaduras son la forma más severa de estrés que el cuerpo puede sufrir; pueden generarse por diferentes agentes térmicos y químicos. CASO CLÍNICO: Paciente de 25 años, con dolor intenso en la región genital de 12 horas de evolución, secundario a la introducción en la vagina de una piedra de alumbre. Se le hicieron múltiples irrigaciones con solución salina al 0.9% sin obtener el resto de la piedra de alumbre. Se le aplicó sulfadiazina de plata en la cavidad vaginal cada 12 horas, óvulos vaginales de ketanserina, miconazol y metronidazol cada 8 horas, ketorolaco por vía oral 10 mg cada 8 horas. Durante su estancia hospitalaria tuvo buena evolución, con disminución de la inflamación en la zona genital, epitelización adecuada. Al tercer día se dio de alta del hospital con cita para valoración a los siete días. CONCLUSIÓN: El tratamiento de las quemaduras en el área genital, por agentes químicos, tiene como piedra angular la identificación del agente causante de la lesión que permita actuar de forma inmediata y evitar las secuelas físicas, sexuales y psicológicas mediante el lavado exhaustivo con solución o agua estéril para remover el agente causal y disminuir que continúe actuando en el sitio afectado.
Abstract BACKGROUND: Burns are the most severe form of stress that the body can suffer; they can be caused by various thermal and chemical agents. CLINICAL CASE: A 25-year-old female patient presented with severe genital pain of 12 hours' duration, secondary to the introduction of an alum stone into the vagina. She underwent several irrigations with 0.9% saline without obtaining the rest of the alum stone. She was given vaginal silver sulfadiazine every 12 hours, vaginal ketanserin, miconazole and metronidazole every 8 hours and oral ketorolac 10 mg every 8 hours. During her stay in hospital, she progressed well, with a decrease in genital inflammation and adequate epithelialisation. She was discharged on the third day with an appointment for a seven-day follow-up. CONCLUSION: The management of genital burns caused by chemical agents is based on the identification of the agent causing the lesion, which allows immediate action and prevents physical, sexual and psychological sequelae by thorough washing with sterile solution or water to remove the causative agent and reduce its continued action in the affected area.
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Objective:To investigate the effects of alum ice nanoemulsion on VEGF and TGF-β1 in hypertrophic scar based on Notch signaling pathway.Methods:Totally 144 SD rats were divided into blank control group, model group, triamcinolone acetonide group and alum ice nanoemulsion low-, medium- and high-dose groups according to random number table method, with 24 rats in each group. Except for the blank control group, the rats in other groups were prepared with deep Ⅱ ° burn models. 24 hours after the successful modeling, the model group was given the same amount of normal saline, the rats in alum ice nanoemulsion low-, medium- and high-dose groups were given 8.15, 6.30 and 32.60 mg/ml alum ice nanoemulsion respectively, and the triamcinolone acetonide group was given triamcinolone acetonide twice a day, 0.2 ml each time, for 35 consecutive days. At 14, 21, 28 and 35 d, the collagen fiber surface density was calculated by VG staining. The protein expressions of vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1), Notch1 and Jagged1 were detected by Western Blot. The expressions of Notch1 mRNA and Jagged1 mRNA were detected by RT-PCR.Results:Compared with model group, triamcinolone acetonide and different doses of alum ice nanoemulsion groups could decrease collagen fiber surface density, protein expressions of VEGF, TGF-β1, Notch1, Jagged1 and mRNA expressions of Notch1, Jagged1 in different degrees ( P<0.05). Compared with the triamcinolone acetonide group, the collagen fiber surface density, protein expressions of VEGF, TGF-β1, Notch1 and Jagged1 and mRNA expressions of Notch1, Jagged1 in the alum ice nanoemulsion medium-dosage group decreased ( P<0.05). Conclusion:Alum ice nanoemulsion can inhibit hypertrophic scar formation, and its mechanism is related to down-regulating Notch signal pathway related molecules Notch1, Jagged1 protein and mRNA levels, and then down-regulating VEGF and TGF-β1 protein expressions.
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Objective:To effectively express the receptor binding domain (RBD) of SARS-CoV-2 spike protein in Pichia pastoris and to evaluate its immunogenicity. Methods:The gene encoding the RBD protein was synthesized and cloned into the pPICZαA plasmid. After linearization, the plasmid was transferred and integrated into the genome of Pichia pastoris. The expressed RBD protein in culture supernatant was analyzed by Western blot and Biolayer interferometry. After screening, a single clone expressing the RBD protein was selected. The high-level expression of RBD protein was achieved by optimizing the fermentation process, including the salt concentration adjusting of the medium and induction condition optimization (pH, temperature and duration). The immunogenicity of the expressed RBD protein was evaluated in a mouse model. Results:A single clone with a high expression level of RBD protein was obtained and named RBD-X33. The expression level of RBD protein in the fermentation supernatant reached up to 240 mg/L after optimization of the induction condition (HBSM medium, pH=6.5±0.3, 22℃ and 120 h). In the mouse experiment, the recombinant RBD protein was formulated with Alum+ CpG dual adjuvant and injected into mice. The binding IgG antibody levels were up to 2.7×10 6 tested by ELISA and the neutralizing antibody levels were up to 726.8 tested by live virus neutralizing antibody assay (prototype). Conclusions:The RBD protein could be efficiently expressed in Pichia pastoris and induce stronger immune response in animals. This study suggested that the recombinant SARS-CoV-2 RBD protein expressed in Pichia pastoris could serve as a candidate antigen in the development of SARS-CoV-2 vaccine.
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ABSTRACT Background Post-operative pain is the major consequence of Ksarasutra, Seton technique employed in Ayurvedic management of Low anal fistula. Surgeons are forced to prescribe Opioids and NSAIDs with pronounced untoward effects. Non pharmacological measures like Balneotherapy are used to improve circulation and relieve spasm in contemporary sciences. Aim To compare the efficacy of Khadira (Acacia catechu) and Sphatika (Potash alum) hot sitzbath with plain hot sitzbath in patients of low anal fistula treated with Ksarasutra. Method The study was single blind, double armed; prospective, randomized control clinical trial in which 30 patients were randomly allocated into two groups of 15 each on the basis of inclusion and exclusion criteria. Group A received hot sitzbath using Acacia catechu and alum infusion twice daily for 21 days. Group B received hot sitzbath using warm water for 21 days. The assessments were made on pain, post-surgical satisfaction burning sensation, tenderness, discharge, constipation, itching and incontinence. Result The disease was prevalent in the 4th decade of life, more in males (86.67%) involved in sedentary work (53.33%) residing in urban domicile, consuming mixed diet (100%). Both groups showed statistically significant improvement in outcome measures like pain, burning sensation, tenderness, discharge, constipation and itching. Conclusion On comparative analysis Acacia and Alum hot sitzbath was more effective in outcomes like pain, surgical satisfaction, burning sensation and discharge. There was no difference in the effect of both interventions with respect to outcome measures like constipation, itching and incontinence.
RESUMO Justificativa A dor pós-operatória é a principal consequência da técnica que utiliza seton de Ksarasutra no tratamento ayurvédico de fístula anal baixa. Os cirurgiões são impelidos a prescrever opiáceos e AINEs que possuem efeitos indesejáveis pronunciados. Medidas não farmacológicas como a balneoterapia são usadas nas ciências contemporâneas para melhorar a circulação e aliviar o espasmo. Objetivo Comparar a eficácia do banho quente de assento com Khadira (Acacia catechu) e Sphatika (Potash alum) com apenas banho de assento em pacientes com fístula anal baixa tratados com Ksarasutra. Método Estudo clínico prospectivo, cego e controlado de dois braços randomizados, no qual 30 pacientes foram alocados aleatoriamente em dois grupos de 15 pacientes cada com base nos critérios de inclusão e exclusão. O Grupo A recebeu banho de assento com Acacia catechu e infusão de alume duas vezes ao dia por 21 dias. O Grupo B recebeu banho de assento com água morna por 21 dias. Os desfechos avaliados foram dor, satisfação pós-cirúrgica, sensação de queimação, sensibilidade, corrimento, constipação, prurido e incontinência. Resultado A doença foi prevalente na 4ª década de vida, mais frequente no sexo masculino (86,67%), nos envolvidos em trabalho sedentário (53,33%), nos residentes em domicílios urbanos e nos que consumiam dieta mista (100%). Ambos os grupos apresentaram melhora estatisticamente significativa na avaliação dos resultados, tais como dor, sensação de queimação, sensibilidade, corrimento, constipação e prurido. Conclusão Na análise comparativa, o banho quente de assento com acácia e alume foi mais eficaz nos desfechos de dor, satisfação cirúrgica, sensação de queimação e corrimento. Não houve diferença em relação ao efeito de ambas as intervenções sobre os desfechos de constipação, prurido e incontinência.
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Humans , Male , Female , Pain, Postoperative , Baths , Rectal Fistula/surgery , Areca , Potassium , Aluminum Sulfate , AcaciaABSTRACT
Background: Radiation cystitis is a well-known complication arising from radiation therapy in pelvic malignancies. Haematuria is usually the main presenting symptom of radiation cystitis, which can range from mild to severe refractory and life-threatening forms. There are various forms of treatment ranging from intravesical instillations like alum, formalin, prostaglandins and pentosan sulphate; hyperbaric oxygen therapy to surgical treatment like cystoscopic fulguration, arterial ligation and cystectomy.Here authors aimed to find the efficacy of 1% alum irrigation in controlling haematuria due to radiation cystitis and to correlate its use with change in clinical status and biochemical parameters.Methods: This retrospective study was done for a period from July 2010 to July 2017 which included all female cervical cancer patients with intractable haematuria due to radiation cystitis. Only patients with normal renal function were included. Irrigation with 0.9% NS, hem coagulase and subsequently 1% alum irrigation were given. Failed cases underwent cystoscopic fulguration.Results: Average age of presentation was 59.79 years. Of the 34 patients 20 (64.51%) patients showed complete response, 4 (12.9%) patients showed partial response and 7 (22.58%) patients had no response to alum irrigation and cystoscopic fulguration were done in these cases. There was no significant change in vital parameters and biochemical parameters except increased prothrombin time (<0.001).Conclusions: No standard of care therapy is presently available for patients with hematuria following radiation cystitis although there is big armamentarium of therapies. Authors found that 1% alum irrigation is safe and efficacious for these patients although normal renal function is a prerequisite.
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Objective: Ammonium alum is a common counterfeit of Alumen,and the processed product of ammonium alum is a common counterfeits of calcined Alumen. This paper aims to establish a method for identifying Alumen,calcined Alumen,ammonium alum and their processed products. Method: The samples were analyzed by scanning electron microscope (SEM) and X ray diffraction (XRD) in this paper. Result: Ammonium alum and Alumen showed obvious changes in morphology after processing. Both Alumen and ammonium alum showed obvious differences in morphology at×250 and×1 000 times microscope. Alumen presented irregular fragments,clear edge corners,smooth surface,scattered irregular small particles,occasional holes and longitudinal edges. Ammonium alum presented irregular clumps,blunt edges,not obvious edges and corners,uneven surface,scattered smaller and round-like particles. The difference in morphology was not obvious at×250 times microscope between Alumen and ammonium alum processed products. While at×1 000 times,the surface of calcined Alumen was uneven with coarse particles; the surface of counterfeit calcined Alumen was flat,and the coarse particle characteristics were not obvious. XRD can be used to rapidly and accurately identify the primary phase of Alumen,calcined Alumen,ammonium alum and ammonium alum processed products:KAl(SO4)2·12H2O,NH4Al(SO4)2·12H2O,KAl(SO4)2,and NH4Al(SO4)2 respectively, with 2θ angle characteristic value of 23,12,22 and 5 respectively for XRD peak. Conclusion: SEM and XRD techniques can be used for the identification of Alumen,calcined Alumen,ammonium alum and their counterfeit products.
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Objective: To provide the new quality control means for Alumen by investigating the elemental differences between calcined Alumen and its counterfeit processed products of ammonium alum, and establishing their characteristic chromatogram. Method: The contents of 22 inorganic elements both in calcined Alumen and processed products of ammonium alum were determined by means of inductively coupled plasma (ICP)-optical emission spectrometer-mass spectrometry (ICP-OES/ICP-MS),SPSS 16.0 was used for cluster analysis (CA) while SIMCA-P 13.0 with t-test and Rank-Sum test was used to identify the differential inorganic elements. In addition, the characteristic spectrum of the inorganic elements for calcined Alumen and counterfeit calcined alumen were established. Result: Calcined Alumen had highest contents of K and Al while counterfeit calcined Alumen has highest contents of Al and Fe;Cr,Sr,and Mn contents in calcined Alumen were relatively higher,while Mn,Ti,and Ga contents in processed products of ammonium alum were relatively higher. The content of K in calcined Alumen was about 205 times of that of counterfeit products. On the contrary,the average contents of Fe,Ti,Mn and Ga in counterfeit products of ammonium alum were much higher than those in calcined Alumen,33,46,38, 27 times, respectively. A total of 18 samples were clustered into two categories in CA:calcined Alumen and processed products of ammonium alum. 18 inorganic elements showed significant difference in contents(PConclusion: This method can be used for quality control of calcined Alumen.
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Objective:To optimize the processing technology of alum by using oven processing combined with central composite design and response surface method to provide basis for processing standardization production of alum. Methods:Alum as the raw ma-terial and on the basis of single factor experiment, the processing temperature, processing time and alum amount were used as the three independent variables factors. A titration method was applied in the content determination of aluminium potassium sulfate in withered alum, and a central composite design and response surface method was used to optimize the processing technology of alum. Results:The optimal processing technology of alum was as follows:the processing temperature was 250℃ with 40-minute processing time and 8 g alum. Conclusion:The optimized alum processing technology by central composite design and response surface method is simple, stable and predictable, which can provide basis for the processing standardization production of alum.
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Objective:To study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs.Methods:BABL/c mice were randomly divided into VLPs experimental group, alum adjuvant experimental group, PBS control group and alum adjuvant control group,the experimental group mice were intramuscular immunization with HBoV1 VP2 VLPs and HBoV1 VP2 VLPs added alum,control group mice were immunization with alum or PBS buffer,then to study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs by cellular and humoral immune strength.Results: Alum adjuvant decreased cellular immune response induced by HBoV1 VP2 VLPs(P<0.001),enhance the HBoV1 VP2 VLPs immuned serum IgG titer(P<0.05)and IgG activity(P<0.01).Conclusion:Alum adjuvant can enhance humoral immune response induced by HBoV1 VP2 VLPs,but weaken cellular immune response induced by HBoV1 VP2 VLPs,when HBoV1 VP2 VLPs used as a prophylactic vaccine it should add alum adjuvant,when used as a therapeutic vaccine,it should not add alum adjuvant.
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Vaccines are some of the most effective tools for the prevention of infectious diseases. Adjuvants are included in vaccines for a variety of reasons: to increase the breadth of response, to lower antigen dose, to overcome limited immune response in some populations, or to enable complex combination vaccines. This study aims to review the safety of licensed vaccine adjuvants and describe their mechanism of action. Potential publications for inclusion were identified through a direct search of PubMed/Medline database. Results of online literature searches were supplemented by relevant papers cited in published studies along with the authors’ knowledge of published studies. To date, there are 5 licensed vaccine adjuvants in US and Europe: Aluminum salts (EU, US), MF59 (EU), AS03 (EU), AS04 (EU, US), and virosomes (EU). AS03 is not available as an adjuvant in other vaccines but included within the US government’s National Stockpile. All vaccines that contain these adjuvants have been proven safe in clinical trials and post-marketing studies, with the exception of the AS03, for which the rare events of narcolepsy have been reported in some countries. Every adjuvant has a complex and often multifactorial immunological mechanism, usually poorly understood in vivo. The safety profile of an adjuvant, including the actual and hypothetical risks, is a critical component that can speed up or impede adjuvant development. The increasing understanding in adjuvant sciences is fundamental to the further development of new adjuvants.
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Vaccines are some of the most effective tools for the prevention of infectious diseases. Adjuvants are included in vaccines for a variety of reasons: to increase the breadth of response, to lower antigen dose, to overcome limited immune response in some populations, or to enable complex combination vaccines. This study aims to review the safety of licensed vaccine adjuvants and describe their mechanism of action. Potential publications for inclusion were identified through a direct search of PubMed/Medline database. Results of online literature searches were supplemented by relevant papers cited in published studies along with the authors’ knowledge of published studies. To date, there are 5 licensed vaccine adjuvants in US and Europe: Aluminum salts (EU, US), MF59 (EU), AS03 (EU), AS04 (EU, US), and virosomes (EU). AS03 is not available as an adjuvant in other vaccines but included within the US government’s National Stockpile. All vaccines that contain these adjuvants have been proven safe in clinical trials and post-marketing studies, with the exception of the AS03, for which the rare events of narcolepsy have been reported in some countries. Every adjuvant has a complex and often multifactorial immunological mechanism, usually poorly understood in vivo. The safety profile of an adjuvant, including the actual and hypothetical risks, is a critical component that can speed up or impede adjuvant development. The increasing understanding in adjuvant sciences is fundamental to the further development of new adjuvants.
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Objective: To evaluate and optimize the origin integration method and process of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine (PRPZA). Methods: Taking the content of calcium oxalate crystal as an index to screen the way of adding ginger and heating method, taking the content of calcium oxalate crystal, the content of water-soluble extracts, and amount of alum residue as the indexes, with the method of orthogonal design and the comprehensive scoring to optimize the processing technology of fresh Pinelliae Rhizoma. Results: The optimized processing method and technology was for per 100 g fresh Pinelliae Rhizoma, plus alum 10 g, mashed ginger 20 g, the common heating to boil after 30 min, soaking for 3 d, and then to 120 ℃ pressure cooking 40 min, rinsing to semi dry, being sliced and dried. Conclusion: The selected origin integration method and technology could not only reduces the content irritant toxic components of calcium oxalate crystal, also avoids the losses of active ingredients, and improve the quality of decoction pieces.
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Potash alum has different traditional application including deodorant and astringent. In order to wide advancement in development of pharmaceutical and cosmetic industries we preferred to work in this field as well as for improvement of safety and efficacy of traditionally used natural products. The aim of the present study was to formulate deodorant potash alum as lotion dosage form and astringent as cream and gel dosage forms. The activity of potash alum against axillaries normal microbiota was tested in vitro using wells diffusion agar method for different concentrations (0.1% to 9%) on Mueller Hinton (MH) agar and the minimum inhibitory concentration (MIC) was determined as 2% and also subculture was done to determine the activity of potash alum as bacteriostatic. Alum powder has wide traditional usage as astringent to prevent bleeding from minor cuts and for its accelerated healing. Therefore, a more appropriate dosage form was designed in a suitable concentration (2% gel) to elongate its astringent activity by its higher retention time due to highly viscous nature of gel. Using suitable excipients, 2% lotion and gel were prepared by dispersion method while emulsification was used for preparation of the potash alum cream of same strength. The above formulations were evaluated by comparing their pH, viscosity, spreadability, contents uniformity and in vitro diffusion. The stability study was carried out at 25°C for 3 months and at 40°C with 75±5% RH for one month. pH, viscosity, spreadability, extrudability and consistency of the products were evaluated at the end of the studies. The results indicated no change in the formulations demonstrating their stable nature as a dosage form and suitability as a commercial product line.
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We report a case of successful treatment for bladder hemorrhage with intravesical alum irrigation. A 60 s woman, who had renal pelvis carcinoma and moderate renal dysfunction（eGFR＝48 mL／min／1.73 m<SUP>2</SUP>）, was hospitalized to our palliative care unit, for pain control of her lumber metastasis. During hospitalization, the patient showed dysuria and a lot of intravesical blood coagulum, which was caused from bladder metastasis. After bladder flushing, continuous bladder irrigation with saline was started. However, when we reduced the irrigation speed, dysuria appeared again. Thus, we started intravesical alum irrigation. Six days after this irrigation has started, we stopped the irrigation because no gross hematuria was seen any more. On the blood test done ninth day after irrigation, serum alminium level was 0.4μg／dL（normal value＜0.8μg／dL）. Three months after discharge from the hospital, the patient died, however, no gross hematuria or anuria were seen up to that time. Serious adverse effects with intracesical alum irrigation were reported in patients with renal dysfunction. However, if the renal dysfunction was moderate, intravesical alum irrigation with reduced doses seem to be safe and effective.
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Potash alum has different traditional application including deodorant and astringent. In order to wide advancement in development of pharmaceutical and cosmetic industries we preferred to work in this field as well as for improvement of safety and efficacy of traditionally used natural products. The aim of the present study was to formulate deodorant potash alum as lotion dosage form and astringent as cream and gel dosage forms. The activity of potash alum against axillaries normal microbiota was tested in vitro using wells diffusion agar method for different concentrations (0.1% to 9%) on Mueller Hinton (MH) agar and the minimum inhibitory concentration (MIC) was determined as 2% and also subculture was done to determine the activity of potash alum as bacteriostatic. Alum powder has wide traditional usage as astringent to prevent bleeding from minor cuts and for its accelerated healing. Therefore, a more appropriate dosage form was designed in a suitable concentration (2% gel) to elongate its astringent activity by its higher retention time due to highly viscous nature of gel. Using suitable excipients, 2% lotion and gel were prepared by dispersion method while emulsification was used for preparation of the potash alum cream of same strength. The above formulations were evaluated by comparing their pH, viscosity, spreadability, contents uniformity and in vitro diffusion. The stability study was carried out at 25°C for 3 months and at 40°C with 75±5% RH for one month. pH, viscosity, spreadability, extrudability and consistency of the products were evaluated at the end of the studies. The results indicated no change in the formulations demonstrating their stable nature as a dosage form and suitability as a commercial product line.
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Objective:To prepare Dermatophagoides farinae (Der f) crude protein to establish BALB/c bronchial asthma model , and to observe the morphology and degranulation of mast cells and detect related cytokines .Methods: Dermatophagoides farinae ( Der f) crude protein were prepared by trituration .30 BALB/c mice were randomly divided into 3 groups:PBS control group (A), asthma model group (B) and Der f crude protein treatment group (C).Group A were treated with PBS(100 μl) all the time, group B and group C were treated with 50 μg Der f crude protein mixed with 50μl alum adjuvant on day 0,day 7 and day 14.On day 28 group A and B were subcutaneous injected with PBS (100 μl) and group C were subcutaneous injected with Der f crude protein (350μg) in PBS(100 μl) at 1-day intervals.One week after the last treatment ,group A,B and C were intranasally challenged with 50 μg Der f crude protein daily for seven days .Twenty-four hours after the last challenge , airway hyper-responsiveness ( AHR) was assessed by using whole-body plethysmography .Two days post challenged , mice were sacrificed and bronchoalveolar lavage fluid ( BALF) was collected.Number of the total cells and eosinophil was determined .Level of IL-4,IL-10 and IFN-γcytokines in the BALF and the su-pernatant of splenocyte culture was assayed by ELISA .Level of Der f specific IgE and histamine in the sera was determined by ELISA . Airway inflammation was analyzed by HE staining .Observation of the morphology and degranulation of mast cells was analyzed by tolui -dine blue staining.Results:Compared with group B,AHR and the lung inflammation in group C were greatly reduced (P<0.01). Numbers of total cells and eosinophils in BALF of group C were significantly lower than that of group B ( P<0.01 ) .Compared with group B, the observation of degranulation of mast cells was insignificant in group C .Compared with group B(IgE:1.905), the level of specific IgE was significantly lower in groups C (IgE:1.278)(P<0.01).The level of IL-4 in BALF of group C was significantly lower than that of group B(P<0.01).Compared with group A and B, the level of IL-10 in BALF was significantly higher in group C (P<0.01) and the level of IFN-γin BALF of group C was significantly higher than that of group A and B (P<0.01).Compared with group B, the level of IL-4 in cultured splenocytes was significantly lower in group C (P<0.01), and the level of IL-10 and IFN-γin cultured splenocytes of group C was significantly higher than that of group B (P<0.01).Compared with group B, the level of histamine in BALF was slightly lower in groups C (P<0.05), and the level of histamine in sera was significantly lower in groups C (P<0.05). Conclusion:The degranulation of mast cells of murine bronchial asthma model was suppressed after the desensitization therapy .
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Aluminum hydroxide (alum) is the most widely used adjuvant in human vaccines. Nevertheless, it is virtually unknown whether alum acts on B cells. In the present study, we explored the direct effect of alum on Ig expression by murine B cells in vitro. LPS-activated mouse spleen B cells were cultured with alum, and the level of isotype-specific Ig secretion, IgG1 secreting cell numbers, and Ig germ-line transcripts (GLT) were measured using ELISA, ELISPOT, and RT-PCR, respectively. Alum consistently enhanced total IgG1 production, numbers of IgG1 secreting cells, and GLTgamma1 expression. These results demonstrate that alum can directly cause IgG1 isotype switching leading to IgG1 production.
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Animals , Humans , Mice , Alum Compounds , Aluminum Hydroxide , B-Lymphocytes , Cell Count , Enzyme-Linked Immunosorbent Assay , Enzyme-Linked Immunospot Assay , Hydroxides , Immunoglobulin Class Switching , Immunoglobulin G , Spleen , VaccinesABSTRACT
Objective To evaluate the immuno-potentiating effects of CpG-ODN plus alum as a composite adjuvant on influenza split virion vaccine.Methods BALB/c mice were immunized with various amounts of 2009 H1N1 influenza split virion vaccine,alone or in combination with CpG-ODN,alum,or both (composite adjuvant).Antigen-specific humoral immune responses were evaluated by ELISA,hemagglutination inhibiting (HI) assay and neutralizing assay.Antigen-specific cellular immune responses were evaluated by ELISPOT assay,intracellular cytokine staining assay and in vivo CTL assay.Results Compared with the control group immunized with antigen alone,a single use of either adjuvant weakly enhanced the humoral immune responses,as indicated by the increase of antigen-specific IgG titers,HI titers and neutralizing titers by 3-6 folds,2-4 folds and 4-8 folds,respectively,after two immunizations.In contrast,the composite adjuvant induced more potent humoral immune responses; the antigen-specific IgG titers,HI titers and neutralizing titers were increased by 23-57 folds,9-20 folds and 16-64 folds,respectively.Consequently,the composite adjuvant achieved antigen-sparing by at least 16 folds.In addition,the composite adjuvant significantly enhanced the antigen-specific cellular immune responses,as revealed by the increase of IFN-γ-secreting CD4+ T cells and the enhancement of CTL activity in immunized mice.Conclusion CpG-ODN plus alum as a composite adjuvant can enhance the immunogenicity of influenza split virion vaccine and achieve the antigen-sparing effect.
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Formalin-killed promastigotes (FKP) of Leishmania major, in combination with Montanide ISA 720 (MISA), BCG or alum were used in vaccination of an inbred murine model against cutaneous leishmaniasis (CL). Significant and specific increases in anti-FKP IgG responses were detected for both alum-FKP and BCG-FKP compared to MISA-FKP (p < 0.001). Significant increases in splenic lymphocyte recall proliferation was obtained in the MISA-FKP vaccinated mice compared to alum-FKP or BCG-FKP vaccinated groups (p < 0.01). The highest interferon-ã responses were observed in the BCG-FKP group followed by the MISA-FKP while the alum-FKP gave the least responses. Significantly reduced lesion sizes were obtained in the MISA-FKP group compared to the BCG/alum adjuvants-FKP vaccinated groups. Although the BCG-FKP group showed the highest IFN-ã responses, it failed to control cutaneous lesions. Significant reductions in parasite numbers were observed in the MISA-FKP and BCG-FKP vaccinated groups (p < 0.001). There was a good correlation between parasite burden and IFN-ã level indicating IFN-ã response as a sensitive parameter of the immune status. In conclusion, MISA-FKP is the most efficacious vaccine formulation against murine cutaneous leishmaniasis.
Promastigotos mortos pela formalina (FKP) de Leishmania major combinados com Montanide ISA 720 (MISA), BCG ou alumen foram usados na vacinação de modelo murino cutâneo de leishmaniose (CL). Aumento significante e específico de resposta IgG anti FKP foram detectados tanto no FKP com alumen como naquele com BCG comparados ao MISA-FKP (p < 0,001). Aumento significante da proliferação esplênica de linfócitos de memória foi obtida nos camundongos vacinados com MISA-FKP quando comparados aos grupos vacinados com alumen-FKP ou BCG-FKP (p < 0,01). As maiores respostas por interferon-gama foram observadas no grupo BCG-FKP seguido pelo MISA-FKP enquanto que o alumen-FKP deu a menor resposta. No grupo MISA-FKP foram obtidas reduções significantes do tamanho das lesões quando comparado aos grupos vacinados com BCG/adjuvante de alumen-FKP. Embora o grupo BCG-FKP tenha mostrado a maior resposta por interferon-gama, não houve controle das lesões cutâneas. Redução significante no número de parasitas foi observada tanto no grupo vacinado com MISA-FKP como no BCG-FKP (p < 0,001). Houve boa correlação entre a carga parasitária e o nível de interferon-gama indicando que a resposta do interferon-gama é parâmetro sensível do estado imunológico. Em conclusão, MISA-FKP é a forma mais eficaz de vacina contra a leishmaniose cutânea murina.