From biospace from noster-it.com Thu Dec 11 01:28:35 2008 From: biospace from noster-it.com (biospace@noster-it.com) Date: Thu Dec 11 12:08:52 2008 Subject: [Immunology] Monitoring Immune Responses In Disease Message-ID: <51a84bbe-bbcd-4808-8830-5c981d2c7fd3@k1g2000prb.googlegroups.com> A recent study published in Clinical Immunology, the official journal of the Clinical Immunology Society (CIS), describes a new method enabling the detection of multiple parameters of single human cells. The report demonstrates the characterization of specific blood cells from an individual with type 1 diabetes, providing information about the role these cells might play in the development of the disease and during therapy. Classification of blood cells, including B and T cells, is important for distinguishing immune responses to pathogens, allergens, or self- antigens in autoimmune diseases. Although various techniques are available to identify cell surface determinants, cytokines and antibodies secreted by blood cells, so far it has not been possible to study multiple secreted proteins while also assigning surface displayed markers to individual living cells. Tonny -------------- Portal to share biological information-data between people http://biospace.ethz.ch From abradbury from taptonschool.co.uk Fri Dec 12 04:36:26 2008 From: abradbury from taptonschool.co.uk (Alan Bradbury) Date: Fri Dec 12 12:04:38 2008 Subject: [Immunology] What causes maternal antibodies to break down in a newborn? Message-ID: I hope somebody can help with this. A recent exam question for A level (16-18 year olds) in the UK had a question asking why passive immunity in newborns is short lived. The answer expected was that a newborn will recognise the maternal antibodies as foreign and have an immune response to them. This just didn't sound right to me. I have tried to do some internet research on the question and it seems that the half-life of maternal antibodies is about 30 - 45 days. But I have not been able to find out what causes the removal of the maternal antibodies. I had assumed that it was just due to the natural turnover and breakdown of the antibodies that the infant might receive from its mother. Am I right, or is the exam answer right? Alan Bradbury (A level Biology teacher, UK). The information contained in this e-mail is confidential and may be privileged. It is intended for the addressee only. If you are not the intended recipient, please notify the sender using the email reply option and then delete it. The contents of this e-mail must not be disclosed or copied without the sender's consent. We cannot accept any responsibility for viruses, so please scan all attachments. From chall from berry.edu Sat Dec 13 13:09:32 2008 From: chall from berry.edu (Chris Hall) Date: Sat Dec 13 15:07:16 2008 Subject: [Immunology] Re: Immuno Digest, Vol 40, Issue 2 In-Reply-To: <200812131703.mBDH3lV03522@net.bio.net> Message-ID: That seems unlikely given that throughout late fetal gestation, antibodies are being passed from mother to fetus, not to mention post-partum transfer through colostrum. The potential for a damaging host vs. graft reaction through immune complex formation would be of concern in removal were through acquired immune mechanisms. Maybe they had in mind innate phagocytosis? I'll be interested in hearing other opinions. > From: > Reply-To: > Date: Sat, 13 Dec 2008 12:03:48 -0500 (EST) > To: > Subject: Immuno Digest, Vol 40, Issue 2 > > Send Immuno mailing list submissions to > immuno@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/immuno > or, via email, send a message with subject or body 'help' to > immuno-request@net.bio.net > > You can reach the person managing the list at > immuno-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Immuno digest..." > > > Today's Topics: > > 1. What causes maternal antibodies to break down in a newborn? > (Alan Bradbury) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 12 Dec 2008 09:36:26 -0000 > From: "Alan Bradbury" > Subject: [Immunology] What causes maternal antibodies to break down in > a newborn? > To: > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > I hope somebody can help with this. > > A recent exam question for A level (16-18 year olds) in the UK had a question > asking why passive immunity in newborns is short lived. The answer expected > was that a newborn will recognise the maternal antibodies as foreign and have > an immune response to them. This just didn't sound right to me. I have tried > to do some internet research on the question and it seems that the half-life > of maternal antibodies is about 30 - 45 days. But I have not been able to find > out what causes the removal of the maternal antibodies. I had assumed that it > was just due to the natural turnover and breakdown of the antibodies that the > infant might receive from its mother. > > Am I right, or is the exam answer right? > > Alan Bradbury (A level Biology teacher, UK). > > The information contained in this e-mail is confidential and may be > privileged. It is intended for the addressee only. > If you are not the intended recipient, please notify the sender using the > email reply option and then delete it. > The contents of this e-mail must not be disclosed or copied without the > sender's consent. > We cannot accept any responsibility for viruses, so please scan all > attachments. > > > ------------------------------ > > _______________________________________________ > Immuno mailing list > Immuno@net.bio.net > http://www.bio.net/biomail/listinfo/immuno > > End of Immuno Digest, Vol 40, Issue 2 > ************************************* From oscar.martinez from syva.es Mon Dec 15 01:42:34 2008 From: oscar.martinez from syva.es (Oscar Martinez Arias) Date: Mon Dec 15 14:50:46 2008 Subject: [Immunology] What causes maternal antibodies to break down in a newborn? In-Reply-To: References: Message-ID: Hi, Alan. The maternal antibodies decrease gradually becuse of their cathabolism, just as the antibodies that are injected into the patients treated with serotherapy. That is the reason for their short live, in both cases. It is not probable that those antibodies cause an immune response. ?scar Mart?nez. -----Mensaje original----- De: immuno-bounces@oat.bio.indiana.edu [mailto:immuno-bounces@oat.bio.indiana.edu] En nombre de Alan Bradbury Enviado el: viernes, 12 de diciembre de 2008 10:36 Para: immuno@magpie.bio.indiana.edu Asunto: [Immunology] What causes maternal antibodies to break down in anewborn? I hope somebody can help with this. A recent exam question for A level (16-18 year olds) in the UK had a question asking why passive immunity in newborns is short lived. The answer expected was that a newborn will recognise the maternal antibodies as foreign and have an immune response to them. This just didn't sound right to me. I have tried to do some internet research on the question and it seems that the half-life of maternal antibodies is about 30 - 45 days. But I have not been able to find out what causes the removal of the maternal antibodies. I had assumed that it was just due to the natural turnover and breakdown of the antibodies that the infant might receive from its mother. Am I right, or is the exam answer right? Alan Bradbury (A level Biology teacher, UK). The information contained in this e-mail is confidential and may be privileged. It is intended for the addressee only. If you are not the intended recipient, please notify the sender using the email reply option and then delete it. The contents of this e-mail must not be disclosed or copied without the sender's consent. We cannot accept any responsibility for viruses, so please scan all attachments. _______________________________________________ Immuno mailing list Immuno@net.bio.net http://www.bio.net/biomail/listinfo/immuno From mrc7 from cam.ac.uk Mon Dec 15 07:59:15 2008 From: mrc7 from cam.ac.uk (Mike Clark) Date: Mon Dec 15 14:50:53 2008 Subject: [Immunology] Re: What causes maternal antibodies to break down in a newborn? References: Message-ID: [Posted and mailed] In message "Alan Bradbury" wrote: > I hope somebody can help with this. > > A recent exam question for A level (16-18 year olds) in the UK had a > question asking why passive immunity in newborns is short lived. The > answer expected was that a newborn will recognise the maternal > antibodies as foreign and have an immune response to them. This just > didn't sound right to me. I have tried to do some internet research on > the question and it seems that the half-life of maternal antibodies is > about 30 - 45 days. But I have not been able to find out what causes > the removal of the maternal antibodies. I had assumed that it was just > due to the natural turnover and breakdown of the antibodies that the > infant might receive from its mother. > > Am I right, or is the exam answer right? > > Alan Bradbury (A level Biology teacher, UK). > I would say that there is little evidence for the exam answer and that the observations are consistent with your answer. Human babies acquire IgG class antibodies from their mother by transport across the placenta in the third trimester of pregnancy. The transport receptor implicated for this is known variously as the Brambell receptor (after the academic who first postulated its existence) or the neonatel Fc receptor (FcRn). Thus young human infants acquire a polyclonal IgG reponse from their mothers. Once the baby is born the only immunoglobulin that the baby can acquire is in the colostrum and later the milk. Milk contains mainly IgM and IgA so this immunoglobulin will mainly pass through, and offer some passive immunity to, the digestive system. In some species such as ruminants and rodents there is little, if any, transplacental transport of IgG and most of the IgG immunoglobulin is transported across the gut by FcRn and is acquired from the colostrum. At present it has not been demonstrated that this is a major mechanism in humans for acquring IgG. Once the baby is born and stops acquring maternal IgG, the immunoglobulin will start to decline for two reasons. [1] Any immunoglobulin that encounters antigen will form an immune complex and be removed from the system via the complement and the Fc receptor mediated processes. [2] Immunoglobulin like any other protein is likely to be destroyed by natural catabolism and so will decline in concentration over time. Interestingly FcRn plays a major role for IgG in reducing the rate of catabolism and thus increasing the half-life of that particular class of antibodies. The examination answer suggests a third possibility. That the infants own anti-globulin response is responsible for removal of maternal IgG. This appears to be invoking an idea related to the Jerne anti-idiotype network hypothesis. That anti-idiotypes (acquired from the mother) will bind to the idiotypes of the infants new B-cells and stimulate them to produce antibody against the mothers antibody. Whilst this is theoretically a possibility I doubt whether it is the major cause of loss of maternal IgG, particularly since the observed decay is mainly consistent with [2] above. Cheers, Mike -- M.R. Clark PhD, Reader in Therapeutic and Molecular Immunology Cambridge University, Department of Pathology Tennis Court Road, Cambridge CB2 1QP Tel +44 (0)1223 333705 Web http://www.path.cam.ac.uk/~mrc7/ From sirgreat76 from hotmail.com Wed Dec 17 06:36:20 2008 From: sirgreat76 from hotmail.com (ali ali) Date: Wed Dec 17 11:01:07 2008 Subject: [Immunology] RE: Immuno Digest, Vol 40, Issue 4 In-Reply-To: <200812161703.mBGH3kV06681@net.bio.net> References: <200812161703.mBGH3kV06681@net.bio.net> Message-ID: Hi All=2C =20 Please guide!=20 =20 I am searching about the role of KIR in Transplantation(Kidney and Liver). = KIR genotyping could be benefical in HLA matched donor and recipient.=20 =20 Raza S. Department of biotechnology University of Karachi. =20 =20 _________________________________________________________________ Suspicious message? There=92s an alert for that.=20 http://windowslive.com/Explore/hotmail?ocid=3DTXT_TAGLM_WL_hotmail_acq_broa= d2_122008= From caroldunlop from yahoo.co.uk Thu Dec 18 05:18:19 2008 From: caroldunlop from yahoo.co.uk (Carol Dunlop) Date: Thu Dec 18 11:05:31 2008 Subject: [Immunology] KIR in Transplantation References: <200812171704.mBHH46V20222@net.bio.net> Message-ID: <63189.13680.qm@web27207.mail.ukl.yahoo.com> Hi Raza: I'm not an expert in the issue but i learnt weeks ago in an Inmunology master class about KIR receptors and alloreactivity in transplantation some interesting things. KIR receptors normally acts as sensors of the cells. They function by binding to class I MHC and when they bind the signaling cascade repress genes that exerts the natural killer function of NK cells. If a cell downregulates the expression of class I MHC (in the case of viral infections for example), then the NK cells don't recognize it and kill that cell. It's known that NK cells pass through a kind of negative selection in bone marrow. There they are exposed to cells that present MHC class I so there is a receptor compatibily selection of the molecules that will be expressed in the surface of the mature and ready to kill NK cells. So in theory I understand that if you select a compatible HLA donor for MHC class I, you are selecting a compatible donor of KIR receptors. However some experiments conclude that in bone marrow transplantation it could be benefical in the 6 months post-transplantation and particullary the graft versus host disease incidence can be diminshed. In Kidney and Liver transplantation it's not allways needed a perfect match for HLA, because these organs seem to be more tolerogenic, specially kidney. I would say that it's not proved that genotyping KIR receptors can help in transplantation of these organs, as they are minor histocompatibility complex and don't have too much variability on the alleles, but you probably will find in the literature this and the oppposite in different studies with different pacients. Unfortunately there's no conclusion as far as i know. Hope to be useful, Carolina Barra Inmunologia Molecular Institut de Biomedicina i Biotecnologia "Vicent Villar i Palasi" Campus UAB-Barcelona-Spain ________________________________ Message: 1 Date: Wed, 17 Dec 2008 11:36:20 +0000 From: ali ali Subject: [Immunology] RE: Immuno Digest, Vol 40, Issue 4 To: , Message-ID: Content-Type: text/plain; charset="Windows-1252" Hi All, Please guide! I am searching about the role of KIR in Transplantation(Kidney and Liver). KIR genotyping could be benefical in HLA matched donor and recipient. Raza S. Department of biotechnology University of Karachi. End of Immuno Digest, Vol 40, Issue 5 ************************************* From webmaster from immunoportal.com Thu Dec 18 04:42:40 2008 From: webmaster from immunoportal.com (webmaster@immunoportal.com) Date: Thu Dec 18 11:06:25 2008 Subject: [Immunology] Immunohistochemistry Message-ID: <699a9055-ae11-4e38-aba5-58dc69a06ac2@s9g2000prm.googlegroups.com> Immunohistochemistry and In Situ Hybridization: A Comprehensive List of Resources on the Internet. Includes a detailed look at immunohistochemistry, immunofluorescence and in situ hybridizations. Discussion board, photogallery, and related links. http://www.immunoportal.com From ashis.rasaily from gmail.com Fri Dec 19 01:06:48 2008 From: ashis.rasaily from gmail.com (ashissr) Date: Fri Dec 19 10:36:36 2008 Subject: [Immunology] COUNCILLING Message-ID: <21086307.post@talk.nabble.com> DEAR ALL, COULD YOU TELL ME THE GOOD PLACES TO DO PHD IN IMMUNOLOGY ASHIS -- View this message in context: http://www.nabble.com/COUNCILLING-tp21086307p21086307.html Sent from the Bio.net - Immuno mailing list archive at Nabble.com. From ashis.rasaily from gmail.com Fri Dec 19 01:01:47 2008 From: ashis.rasaily from gmail.com (ashis.rasaily@gmail.com) Date: Sun Dec 21 14:31:06 2008 Subject: [Immunology] NEED HELP Message-ID: <001485f3be6258e4ae045e600b0b@google.com> THANK YOU FOR YOUR SUGESSTION LIT LI CHEING.I NOW HAVE A PROBLEM REGARDING THE FICOL PROCESS.ie FOR MY PROJECT IMMUNOMODULATORY EFFECTS OF ALOR VERA EXTRACT ON HUMAN PBMCS,I GET THE BLOOD[ WITH EDTA] AND AFTER 15 MINUTES I USE IT FOR FICOL GRADIENT SEDIMENTATION. I DILUTE THE BLOOD WITH EQUAL VOL OF PBS BUFFER TO AVOID QUICK CLUMPING WHICH USED TO EARLIER HAPPEN IN THE NEXT STEP ie WHEN I ADD FICOL. I USUALLY HAVE, 2, 14 ml TUBES OF THIS FICOL-BLOOD SO THE TIME TAKEN FOR ME TO CENTRIFUGE AFTER I HAVE FILLED THE BOTTLE IS APPROX 5-6 MINUTES. I CENTRIFUGE IT IN A SWINGING BUCKET CENTRIFUGE FOR ABOUT 20-25 MINUTES AT A SPEED OF ABOUT 3000 rpm. I THEN GET MY BUFFY COAT. I NOW SEPARATE MY BUFFY COAT USING A PIPETTE.i .e I FIRST REMOVE THE SERUM/PLASMA ON THE VERY TOP FOLLOWED BY THE BUFFY COAT.I WOULD NOW LIKE TO SPECIFY THAT ALONG WITH THE BUFFY COAT I REMOVE EVEN SOME PARTS OF FICOL UNDERNEATH IT AND SOME PARTS OF THE SERUM ABOVE ONLY TO OPTIMISE THE AMOUNT OF THE BUFFY COAT.I THEN ADD EQUAL VOL OF PBS BUFFER TO "WASH" THE BUFFY COAT OR IN IN OTHER TERMS TO PRECIPITATE THE BUFFY COAT."NOW COMES THE PROBLEM" I OFTEN GET A "RED SPEC" SOMETIMES BIG AND SOMETIMES SMALL, OVER MY BUFFY COAT.NOW WHATS THAT? PS:AT THIS POINT I MUST SAY THAT MY CENTRIFUGE IS NOT THAT FINE ie IT HAS ITS JERKS AS IT ROTATES BUT COULD THE "RBC" WHICH I KNOW IS AT THE TOP OF YOUR MIND GO AGAINST THE GRADIENT AT A SPEED OF 3000 RPM? I AM REALLY LOOKING FORWARD FOR YOUR ANSWER AND SHOULD I GO FORWARD WITH MY EXPERIMENT OF SEEKING THE ACTION OF ALOE EXTRACTS ON THE "PBMCS" IF I GET THE RED SPEC? AM REALLY LOOKING FORWARD FOR YOUR REPLY.WISHING YOU "ALL", THE BEST IN YOUR WORKS ASHIS From sushmamanral from gmail.com Tue Dec 23 10:45:35 2008 From: sushmamanral from gmail.com (sushmamanral@gmail.com) Date: Tue Dec 23 14:25:35 2008 Subject: [Immunology] Re: NEED HELP References: Message-ID: On Dec 19, 11:01?am, ashis.rasa...@gmail.com wrote: > THANK YOU FOR YOUR SUGESSTION LIT LI CHEING.I NOW HAVE A PROBLEM REGARDING ? > THE FICOL PROCESS.ie FOR MY PROJECT IMMUNOMODULATORY EFFECTS OF ALOR VERA ? > EXTRACT ON HUMAN PBMCS,I GET THE BLOOD[ WITH EDTA] AND AFTER 15 MINUTES I ? > USE IT FOR FICOL GRADIENT SEDIMENTATION. I DILUTE THE BLOOD WITH EQUAL VOL ? > OF PBS BUFFER TO AVOID QUICK CLUMPING WHICH USED TO EARLIER HAPPEN IN THE ? > NEXT STEP ie WHEN I ADD FICOL. > I USUALLY HAVE, 2, 14 ml TUBES OF THIS FICOL-BLOOD SO THE TIME TAKEN FOR ME ? > TO CENTRIFUGE AFTER I HAVE FILLED THE BOTTLE IS APPROX 5-6 MINUTES. I ? > CENTRIFUGE IT IN A SWINGING BUCKET CENTRIFUGE FOR ABOUT 20-25 MINUTES AT A ? > SPEED OF ABOUT 3000 rpm. I THEN GET MY BUFFY COAT. > I NOW SEPARATE MY BUFFY COAT USING A PIPETTE.i .e I FIRST REMOVE THE ? > SERUM/PLASMA ON THE VERY TOP FOLLOWED BY THE BUFFY COAT.I WOULD NOW LIKE TO ? > SPECIFY THAT ALONG WITH THE BUFFY COAT I REMOVE EVEN SOME PARTS OF FICOL ? > UNDERNEATH IT AND SOME PARTS OF THE SERUM ABOVE ONLY TO OPTIMISE THE AMOUNT ? > OF THE BUFFY COAT.I THEN ADD EQUAL VOL OF PBS BUFFER TO "WASH" THE BUFFY ? > COAT OR IN IN OTHER TERMS TO PRECIPITATE THE BUFFY COAT."NOW COMES THE ? > PROBLEM" I OFTEN GET A "RED SPEC" SOMETIMES BIG AND SOMETIMES SMALL, OVER ? > MY BUFFY COAT.NOW WHATS THAT? > PS:AT THIS POINT I MUST SAY THAT MY CENTRIFUGE IS NOT THAT FINE ie IT HAS ? > ITS JERKS AS IT ROTATES BUT COULD THE "RBC" WHICH I KNOW IS AT THE TOP OF ? > YOUR MIND GO AGAINST THE GRADIENT AT A SPEED OF 3000 RPM? I AM REALLY ? > LOOKING FORWARD FOR YOUR ANSWER AND SHOULD I GO FORWARD WITH MY EXPERIMENT ? > OF SEEKING THE ACTION OF ALOE EXTRACTS ON THE "PBMCS" IF I GET THE RED ? > SPEC? AM REALLY LOOKING FORWARD FOR YOUR REPLY.WISHING YOU "ALL", THE BEST ? > IN YOUR WORKS ASHIS hi i m new to this site n happened to just read ur problem actually i m also doing work on pbmcs or rather to say on lymphocytes the red specs u get may be RBCs u can give osmotic shock to remove this add 0.2%NaCl to the pellet wait for 30 secs and then add an equal volume of 0.16% NaCl dropwise to it and then centrifuge give a wash after that so as to remove any NaCl. hoping this may solve ur problem From sushmamanral from gmail.com Tue Dec 23 10:49:49 2008 From: sushmamanral from gmail.com (sushmamanral@gmail.com) Date: Tue Dec 23 14:25:42 2008 Subject: [Immunology] density of platelets Message-ID: <329a08a4-e2a5-498e-8854-45ea9ae3e299@w24g2000prd.googlegroups.com> hi actually i m separating lymphocytes by using histopaque 1.077 but when i do differential counting i get platelets also. is there any way i can remove these platelets? and kindly tell me whats the density of lymphocytes and platelets, and their diameters too. From de_gloor from bluewin.ch Tue Dec 23 11:52:48 2008 From: de_gloor from bluewin.ch (Denise Gloor) Date: Tue Dec 23 14:25:47 2008 Subject: [Immunology] Re: What causes maternal antibodies to break down in a newborn? References: Message-ID: <495117bf$1@maser.urz.unibas.ch> Test "Mike Clark" schrieb im Newsbeitrag news:b379120e50.mrc7offline@mrc7acorn1.path.cam.ac.uk... > [Posted and mailed] > > In message > "Alan Bradbury" wrote: > >> I hope somebody can help with this. >> >> A recent exam question for A level (16-18 year olds) in the UK had a >> question asking why passive immunity in newborns is short lived. The >> answer expected was that a newborn will recognise the maternal >> antibodies as foreign and have an immune response to them. This just >> didn't sound right to me. I have tried to do some internet research on >> the question and it seems that the half-life of maternal antibodies is >> about 30 - 45 days. But I have not been able to find out what causes >> the removal of the maternal antibodies. I had assumed that it was just >> due to the natural turnover and breakdown of the antibodies that the >> infant might receive from its mother. >> >> Am I right, or is the exam answer right? >> >> Alan Bradbury (A level Biology teacher, UK). >> > > I would say that there is little evidence for the exam answer and that > the observations are consistent with your answer. > > Human babies acquire IgG class antibodies from their mother by transport > across the placenta in the third trimester of pregnancy. The > transport receptor implicated for this is known variously as the > Brambell receptor (after the academic who first postulated its > existence) or the neonatel Fc receptor (FcRn). Thus young human infants > acquire a polyclonal IgG reponse from their mothers. > > Once the baby is born the only immunoglobulin that the baby can acquire > is in the colostrum and later the milk. Milk contains mainly IgM and IgA > so this immunoglobulin will mainly pass through, and offer some passive > immunity to, the digestive system. In some species such as ruminants and > rodents there is little, if any, transplacental transport of IgG and > most of the IgG immunoglobulin is transported across the gut by FcRn and > is acquired from the colostrum. At present it has not been demonstrated > that this is a major mechanism in humans for acquring IgG. > > Once the baby is born and stops acquring maternal IgG, the > immunoglobulin will start to decline for two reasons. [1] Any > immunoglobulin that encounters antigen will form an immune complex and > be removed from the system via the complement and the Fc receptor > mediated processes. [2] Immunoglobulin like any other protein is likely > to be destroyed by natural catabolism and so will decline in > concentration over time. Interestingly FcRn plays a major role for IgG > in reducing the rate of catabolism and thus increasing the half-life of > that particular class of antibodies. > > The examination answer suggests a third possibility. That the infants > own anti-globulin response is responsible for removal of maternal IgG. > This appears to be invoking an idea related to the Jerne anti-idiotype > network hypothesis. That anti-idiotypes (acquired from the mother) will > bind to the idiotypes of the infants new B-cells and stimulate them to > produce antibody against the mothers antibody. Whilst this is > theoretically a possibility I doubt whether it is the major cause of > loss of maternal IgG, particularly since the observed decay is mainly > consistent with [2] above. > > Cheers, > > Mike > -- > M.R. Clark PhD, Reader in Therapeutic and Molecular Immunology > Cambridge University, Department of Pathology > Tennis Court Road, Cambridge CB2 1QP > Tel +44 (0)1223 333705 Web http://www.path.cam.ac.uk/~mrc7/ From ashis.rasaily from gmail.com Thu Dec 25 00:54:05 2008 From: ashis.rasaily from gmail.com (ashissr) Date: Thu Dec 25 16:00:53 2008 Subject: [Immunology] NEED HELP In-Reply-To: References: <001485f3be6258e4ae045e600b0b@google.com> Message-ID: <21165596.post@talk.nabble.com> sushmamanral wrote: > > On Dec 19, 11:01?am, ashis.rasa...@gmail.com wrote: >> THANK YOU FOR YOUR SUGESSTION LIT LI CHEING.I NOW HAVE A PROBLEM >> REGARDING ? >> THE FICOL PROCESS.ie FOR MY PROJECT IMMUNOMODULATORY EFFECTS OF ALOR VERA >> ? >> EXTRACT ON HUMAN PBMCS,I GET THE BLOOD[ WITH EDTA] AND AFTER 15 MINUTES I >> ? >> USE IT FOR FICOL GRADIENT SEDIMENTATION. I DILUTE THE BLOOD WITH EQUAL >> VOL ? >> OF PBS BUFFER TO AVOID QUICK CLUMPING WHICH USED TO EARLIER HAPPEN IN THE >> ? >> NEXT STEP ie WHEN I ADD FICOL. >> I USUALLY HAVE, 2, 14 ml TUBES OF THIS FICOL-BLOOD SO THE TIME TAKEN FOR >> ME ? >> TO CENTRIFUGE AFTER I HAVE FILLED THE BOTTLE IS APPROX 5-6 MINUTES. I ? >> CENTRIFUGE IT IN A SWINGING BUCKET CENTRIFUGE FOR ABOUT 20-25 MINUTES AT >> A ? >> SPEED OF ABOUT 3000 rpm. I THEN GET MY BUFFY COAT. >> I NOW SEPARATE MY BUFFY COAT USING A PIPETTE.i .e I FIRST REMOVE THE ? >> SERUM/PLASMA ON THE VERY TOP FOLLOWED BY THE BUFFY COAT.I WOULD NOW LIKE >> TO ? >> SPECIFY THAT ALONG WITH THE BUFFY COAT I REMOVE EVEN SOME PARTS OF FICOL >> ? >> UNDERNEATH IT AND SOME PARTS OF THE SERUM ABOVE ONLY TO OPTIMISE THE >> AMOUNT ? >> OF THE BUFFY COAT.I THEN ADD EQUAL VOL OF PBS BUFFER TO "WASH" THE BUFFY >> ? >> COAT OR IN IN OTHER TERMS TO PRECIPITATE THE BUFFY COAT."NOW COMES THE ? >> PROBLEM" I OFTEN GET A "RED SPEC" SOMETIMES BIG AND SOMETIMES SMALL, OVER >> ? >> MY BUFFY COAT.NOW WHATS THAT? >> PS:AT THIS POINT I MUST SAY THAT MY CENTRIFUGE IS NOT THAT FINE ie IT HAS >> ? >> ITS JERKS AS IT ROTATES BUT COULD THE "RBC" WHICH I KNOW IS AT THE TOP OF >> ? >> YOUR MIND GO AGAINST THE GRADIENT AT A SPEED OF 3000 RPM? I AM REALLY ? >> LOOKING FORWARD FOR YOUR ANSWER AND SHOULD I GO FORWARD WITH MY >> EXPERIMENT ? >> OF SEEKING THE ACTION OF ALOE EXTRACTS ON THE "PBMCS" IF I GET THE RED ? >> SPEC? AM REALLY LOOKING FORWARD FOR YOUR REPLY.WISHING YOU "ALL", THE >> BEST ? >> IN YOUR WORKS ASHIS > > hi > i m new to this site n happened to just read ur problem > actually i m also doing work on pbmcs or rather to say on lymphocytes > the red specs u get may be RBCs > u can give osmotic shock to remove this > add 0.2%NaCl to the pellet wait for 30 secs and then add an equal > volume of 0.16% NaCl dropwise to it and then centrifuge > give a wash after that so as to remove any NaCl. > hoping this may solve ur problem > _______________________________________________ > Immuno mailing list > Immuno@net.bio.net > http://www.bio.net/biomail/listinfo/immuno > > thank you I'll surely implement that and get back to you. ashis -- View this message in context: http://www.nabble.com/NEED-HELP-tp21118307p21165596.html Sent from the Bio.net - Immuno mailing list archive at Nabble.com. From biospace from noster-it.com Sat Dec 27 01:05:39 2008 From: biospace from noster-it.com (biospace) Date: Sat Dec 27 15:00:50 2008 Subject: [Immunology] Naturally Occurring 'Protective RNA' Used To Develop New Antiviral Against Influenza Message-ID: Researchers from the University of Warwick, Coventry, United Kingdom have developed a new antiviral using naturally occurring influenza virus "protecting virus" that may defend against any influenza A virus in any animal host. Human influenza virus A is not only responsible for seasonal disease in humans, it is also the cause of worldwide pandemics of which the last three resulted in millions of deaths all over the globe. Currently, live and killed vaccines countering specific strains of the flu are available, however a vaccine for a new pandemic strain would take months to develop. Also, increased resistance to antivirals currently on the market has emphasized the need for a new effective prophylactic and therapeutic treatment method. The "protecting virus" contains an altered gene that makes it harmless and prevents it from reproducing in a cell. If another influenza virus invades the cell it still remains harmless, but rapidly reproduces and prevents infection by literally crowding out the new influenza strain. Tonny -------------- More bio-med news & videos Portal to share biological information-data between people http://biospace.ethz.ch