Science Buddies: "Ask an Expert"

Hi,
I want to do a project on psoriasis this year.
I am interested in microbiology projects, especially ones related to skin problems.
My cousin has atopic eczema, and after researching eczema I came across psoriasis.
Psoriasis is caused when there are too many T-cells producing too many skin cells causing red patches on the skin. I read online about some Chinese oriental medicines with the ability to regulate the amount of T-cells. My father knows about Chinese oriental medicine, and I am trying to learn about which herbs I can test.

I want to test the effectiveness of one/two Chinese oriental medicines on how they affect T-cells and connect that to an effective preventative measure against psoriasis.

Questions I have:
1) How would I conduct this experiment, would I need a lab?
2) If I need a lab/mentor, how would I find one?
3) What is an effective way to test this?
-I don't know if I can use human cells/animal cells without a lab
-Is there a method I can use at home?

4) Is there a way I could adjust my experiment to improve it?
-Measure different things
-Different project all together? (this may not feasible...?)

I would appreciate any advice!

Thank you.

Lauren

Hi Lauren,

That's a great project, to investigate Chinese herbal medicines as a remedy for psoriasis. We had a PhD student in our lab who studied the effects of the herb Curcuma longa from which the spice turmeric is obtained on the immune system. She used human blood cells that can be grown and treated in the lab. Unfortuantely, you can't do that at home. You would need to work in a lab under supervision with people who know how to culture and work with human cells. It is not difficult to do these experiments but you do need a biosafety cabinet, CO2 incubator, centrifuge and, of course, access to a source of human T cells or other types of immune cells such as dendritic. You could also culture human skin cells and expose them to the T cells that had been treated with the Chinese herbal extract and measure some change in activity.

If you live reasonably close to a large university, especially one that has a medical school that does grant-funded research, you could search the faculty list for publications. See if there are any people who are doing immunology research using cultured cells, read some of their papers if you can get them (i can help you) then call them and tell them that you have a project in mind to study the efficacy of Chinese herbal medicines on the immune responses of the skin. You will have to do some reading or watch some youtube videos about how the immune system works so you can talk about it intelligently and respond to questions. Working scientists are happy to take in students if they can demonstrate that they are serious and will work hard.

This is a highly interesting project, but it is going to be difficult and take a lot of time since the experiments are really at a college or even graduate level. I don't know how much time you have before your project is due. You would need to set up a strict timetable and be prepared to spend several hours a week in the lab. Usually the high school students we have in the lab come in over the summer when they aren't in school.

Please let me know more about your plans. The only other possibility i can think of is to recruit your friends and ask them if they have used herbal skin treatments, and if so, how well they worked compared to conventional creams and lotions. I'm sure there's lots of information online about Chinese medicine, but there may not be the type of clinical trials like big pharmaceutical companies do for candidate drugs. Based on the existing evidence for efficacy in treating psoriasis, you could choose a group of herbs and see if there are any common over-the-counter preparations containing them. Do you have any Chinese friends? Ask them!

Good luck and do keep us posted about your plans and progress.

Best wishes,

Sybee

Hi Sybee,
Thank you for you reply.
I am constructing a list of professors from immunology, microbiology, and dermatology departments that have done some research on my topics. I am not sure how I should contact them, and how I would introduce my project to them. Tomorrow, I will be going to an Ask-a-Scientist Night sponsored by my district, where scientists and teachers come to help students with their projects. I will hopefully find some more answers, and hopefully an interested mentor.

I am going to continue to expand my basic knowledge about the immune system and psoriasis. I am trying to analyze the information in a couple papers I have attempted to read, but I feel that my background knowledge is not strong enough yet to fully understand the findings.
I have approximately 3-4 months to complete this project, and am willing to devote several hours a week in order to complete it.
I talked with my father and he suggested for me to use one/two of the following herbs, Glycyrrhiza uralensis (Liquorice), Smilax glabra (Glabrous greenbrier), and Carthamus tinctorius (Safflower). These are some medicines that he and his colleagues often use in treating psoriasis at his school and clinic. I know for a fact I will be able to get this ingredients if I continue this experiment.

With my experiment (If I find a mentor and lab), what would I be measuring? I read on another "science buddies" forum post about psoriasis that a student grew t-cells in tissue culture and measured inflammatory cytokines like TNF-alpha, IL-1beta, and IFN gamma. Would I look at the behaviors of the cells and how it has changed?

I am fairly new to this field, and am open to learning as much about it as I can. I really appreciate all your guidance and advice!


Thank you.

Lauren

Hi Lauren,

You are doing great! I am so happy that you are making good progress in finding a lab to work in and a mentor. Most scientists like to help young students because they remember when they were in your position and needed guidance and a place to work! So, don't have any hesitation to approach someone with your proposal. Just make sure you have done your homework and know that the work you propose fits with what their lab has been doing. Go to PubMed and look up their publications over the past five years or so.

Our lab does research on allergy and asthma--inflammation in the airways--so i'm not that familiar with psoriasis, but i do understand pretty well how the immune system works and can answer your questions. You have enough time to do a good project, if you can find a lab that is already set up to do experiments with immune cells such as T cells and dendritic cells or Langerhans cells in the skin. You are right that one way to measure activity of immune system cells is to quantitate their output of specific proteins, called cytokines--the interleukins, TGF-beta, TNF-alpha, etc. You will have to look up and become familiar with the enzyme-linked immunosorbent assay, abbreviated ELISA [pronounced like 'Eliza'] and flow cytometry. These are standard methods for measuring cytokines. The other thing you need to be familiar with is how to culture cells. There are many, many tutorials online that will teach you about these methods. Check YouTube for some videos or slide presentations.

I know this is a lot to learn in a short time, but it is really interesting and is what many scientists are working on now. The immune system affects everything in your body from digestion to the brain and when it misbehaves, we can suffer.

Good luck finding a lab and people to help you. We'll be here to assist!

Sybee

Hi Lauren,

SciB has given you a lot of great advice, and you are doing an amazing job doing some research on your topic of interest and actively trying to find a mentor to help you! If I am understanding correctly, one speculation of how psoriasis is caused is that T cells are activated when they're not supposed to, and the cytokines they release and inflammation that results are what trigger rapid production of skin cells. Some ideas you may like to consider as you develop your project more are:

1. Are you just going to look at the responses of the T cells to the herbal medicines in vitro? It would definitely be cool to see if treatment of T cells with the herbal medicines can trigger a different cytokine secretion profile, but that would not necessarily mean that it has some sort of effect on the development of psoriasis. You would have to try to find a way to relate this to the skin. For example, is there a way that you can co-culture skin cells and T cells, and then treat with the herbal medicines to see how the T cells may affect the proliferation of the skin cells? I'm not an expert on working with skin cells and immune cells, so this is something you should definitely ask your mentor.

2. If you were to work in a lab, have you considered using animal models of psoriasis to see how effective your herbal medicines are in treating this disease condition? This is definitely something you can ask your mentor as well if there is something like that available. It will be cool if you would be able to look at an actual disease setting, and see how the cytokine profiles of the T cells as well as the proliferation status of the skin cells are affected in vivo.

Hope this helped in some way!

Connie

Hi Sybee,
I found a professor who said he would "be happy to discuss your project and guide you".
However, he is out traveling until after thanksgiving. My project is due at the end of January, will I be able to complete this project within that amount of time?
I took notes on the ELISA method (http://www.bio.davidson.edu/genomics/method/ELISA.html I used this website) , and am starting to gain a better understanding of how I would complete this project if I get a lab. I just submitted a form to my county to see if I will get approval in order to use these materials.
Would I need to use a liquid form of my medicines in order to consistently treat the cells? How would I extract a liquid? Is there any specific method you might advise me to use?
My Project Title right now is : The Efficacy of Chinese Herbal Medicines on the Production of Inflammatory Cytokines in Th17 Cells in the Treatment of Psoriasis

Thank you for your advice. I will post any questions I have.

Lauren

Hi Connie,
Thank you for your advice.
Psoriasis is like what you have stated. The skin over-generates Tcells that produce more cells than it can shed off which results in red, flaky patches on the patient's skin.
1. I read online that specific cytokines like IFN-gamma, IL-2 are found in psoriasis. That is why I decided to look at the immune responses of Th17 cells.
I read this article about an experiment done on Th17 cells, and it looks like they just measured the efficacy of how this herb inhibits the growth of Th17 cells.
http://www.webmd.com/rheumatoid-arthrit ... e-diseases
Would it be more efficient to measure the amount of T-cells in skin cells before and after treatment?
Or would observing specific psoriasis related cytokines be an effective experiment?
I think the former would be more of a preventative measure towards inflammation, and the latter would be more of a therapeutic remedy for patients.
What would you suggest?

2. I will definitely ask my mentor that. Unfortunately, he is out of town until after Thanksgiving. Would you suggest for me to find a new mentor? Or wait until he comes back? (My project is due in late January). I think looking at an actual disease setting would be very interesting and an effective way to connect the treatment to the disease.

Thank you very much!

Lauren

Hi Lauren,

Your project has the makings of a real winner, but it is going to be technically very demanding. You really should be starting NOW because you need to learn a lot of methods in a short time in order to get some meaningful data by the end of January. Our whole university shuts down from before Christmas through New Years, so you have to figure that during that period, your time in your lab may also be limited.

Is there no one else in the lab that you had planned to work in that can show you how to isolate, characterize and culture Th17 cells? You don’t need a professor to teach you these methods. A staff technician or graduate student can show you how it is done and watch you while you try it. Ask your mentor if you can possibly get started now. A whole month is a long time to wait when you have so little time. Otherwise, you should find another lab that is doing immune cell research and has the necessary methods and equipment AND where someone is available right now to teach you and help you get going.

As I understand your project you are hypothesizing that Th17 cells are one of the causes of psoriasis and that it is possible to inhibit the activity of these cells with certain Chinese herbal medicines. There is ample evidence that the IL-17/IL-23 pathway that is active in Th17 cells contributes to a variety of immune system diseases including arthritis and psoriasis. Pharmaceutical companies are testing drugs in clinical trials to determine their ability to inhibit Th17 cells or the IL-17/IL-23 pathway, so your experiments are right at the forefront of current research.

While it would be highly interesting to test your medicines in an animal model of psoriasis as Connie suggested, that would be a long-term project unless you were lucky enough to find a lab that was doing these experiments already and that would allow you to work with them to test your herbal preparations.

When we do a project like yours, we usually start out testing on cultured cells before moving on to animal experiments so we have a good idea about toxicity, dosing and effects on the cytokine pathways in the cells. I would suggest that you start by learning how to isolate, culture and characterize Th17 cells. These immune system cells come from what are known as naïve [because they have never been exposed to antigen] T cells that are of a type called CD4+ for a specific protein marker that they carry. When CD4+ T cells are exposed to certain cytokines including transforming growth factor beta [TGFb], interleukin 1 beta [IL-1b] and IL-6, they undergo a change in gene expression that causes them to produce IL-17 and a cell surface receptor for IL-23. When this receptor binds to IL-23 that is produced by T helper cells [another type of immune cell] the CD4+ T cells are converted into Th17 cells. If I am being too technical, please ask me questions and I will try to clarify. Immunology is a complex subject but it is utterly fascinating and the immune system affects everything in your body.

You can isolate CD4+ T lymphocytes from human blood that your mentor can get from a blood bank. The blood is tested for HIV, hepatitis and other pathogens so it is safe, but you still need to handle it under what are called biosafety level 2 conditions. Your mentor can explain how to do this.

Once you get a population of Th17 cells growing in culture and you have shown that they produce IL-17, then you can test a range of doses of your Chinese medicine on the cells. You asked about the form of the medicine and yes, you will need to make a water extract of it and filter-sterilize it before exposing the cells to it. Your father should be able to help you prepare an active extract. Your first experiment will be to determine the toxicity of the preparation on your Th17 cells. You do this by making several dilutions of your medicine—1:5, 1:10, 1:50, 1:100, etc. Then you add a certain volume of the diluted preparation to your cell culture medium and allow the cells to continue growing for a certain length of time [this time is one of the variables you will have to determine]. As a control, you incubate some of the cells with just the vehicle you used to make the herbal extract. There are several ways to measure toxicity and how you do it will depend on how the lab that you work in does it. You can google ‘cell viability assays’ to find out more about this.

After you have tested various doses of your medicine and found a range of amounts that do not kill the cells or stop them from growing, then you can measure the effect of the medicine on the production of IL-17. I’m not sure which method you would use to activate the cells, but your mentors can help you there. What you are looking for is inhibition of IL-17 production by the herbal medicine. You will probably measure IL-17 by ELISA, a straightforward spectrophotometric method that is commonly used in biomedical science labs. Activated Th17 cells also produce IL-22 and IL-26.

Phew! I think I’ll stop there. I’ve given you a lot of suggestions, so after you have thought about it, repost to this thread and let me know your plans. If your mentors are immunologists, they can answer all your questions, but we will always be here to help you with the details.

All the best,

Sybee

Thank you for your advice Sybee.
With all the information I've been able to picture exactly what I would do to test this experiment.
Based on your advice, I tried to come up with an simple experimental design.
"Firstly, I will need to test the toxicity of the Chinese medicines on the Th17 cells. After making a water extract and filter-sterilizing it, I want to test several dilutions of the medicines to see a range of amounts that don't kill the cells or stop them from growing.
Then, I want to measure the effect of the medicine on the production of IL-17. (present in the Th17 cells that cause psoriasis + psoriatic arthritis). I do not know what method I can use to activate the cells, but I will be looking for the inhibition of IL-17 production by the herbal medicine possibly using the ELISA method."

I emailed the professor and he said to only contact him after Thanksgiving. He also said that he has not completely agreed to let me perform this experiment in his lab (until he meets me in person) and says that the reagents needed for this experiment are very expensive.
I am very passionate about this project and my interest continues to grow with the more research I do. This is my first time trying to find a mentor, and I am trying not to feel discouraged, but it's difficult.
I sent out a couple more emails out to some more professors and I am hoping for a positive response.

If I do not find a mentor in a couple weeks, my science teacher advised me to move on to a new project.

Thank you for your advice! I really appreciate everything!

Lauren

Hi Lauren,

You are doing a great job and working like a real scientist to get your project going! Finding a lab to work in IS difficult, but keep trying. Yes your experiments are expensive, but so is everything in biomedical research. That’s why we have to struggle so hard to get funding. I spend one-third of my time writing federal and state grants to get money to do biomedical research and only one grant in six is successful!

I can tell you are getting excited about immunology and the more you find out about it the more interesting it becomes. Keep reading about psoriasis and the new potential treatments based on what we are discovering in the lab. Learn to use the scientific jargon correctly and make sure what you learn is correct. Pay special attention to the methods used in the lab to get good data. I can answer your questions about techniques. When you talk to a prospective mentor, you want them to see how serious you are about this project and that you are competent to do it.

Do you happen to live within a reasonable distance from a college of medicine? I have found that MDs are sometimes more interested in doing medical research projects than academic PhD scientists. They usually have connections with clinical research scientists in whose labs their medical interns can do research projects. This may be another way that you could find a place to do your experiments. People suffering from psoriasis are looking for good new treatments and learning about the role of Th17 cells in the pathology is one good way to develop a new drug.

One other possibility for a mentor is with a biomedical, pharmaceutical or drug development company. Companies do take interns to work on projects in their labs, but they usually want college-level students. Can’t hurt to ask, however, if there are any companies like that in your area.

Even if you don’t get to do this specific project right now, you will have gained a lot of knowledge and be much better prepared when opportunity comes along later. Think about some immunology-related projects you can do now as a back up and talk to your teachers about what resources they have if you can’t get into a university lab. We will continue to help with whatever you decide.

All the best,

Sybee

Hi Sybee
I hope you've been well. Also, I hope you had a wonderful thanksgiving. Sorry for my late posts I've been busy trying to contact my mentor.
I am meeting with him later this week do you know if you could give me some advice on how I should prepare in meeting him?
What should I expect and should I dress formally? I'm really nervous and know that this interview will be the deciding factor to determine if he will allow me to use his laboratory.


Thank you for all your help! I really appreciate it.

Hi Lauren,

Sorry i missed your post. I wish they would send me a text or an email when i get a reply from one of the students i am working with so i know. All the posts appear together and i didn't see yours until I went way down the list.

I have confidence in you and I am sure you did well talking to the professor. They are looking for students who know something about the science and are willing to put in the time to learn the lab methods to do the experiments to answer an important question. I know you are that kind of person and I would have given you a letter of recommendation if that had been possible!

So, tell me what happened and whether you are going to be able to work in the lab. We will be here to continue your virtual mentoring. Don't be afraid to ask any question. If you don't understand something, just say so. None of us was born with this knowledge and there's a ton of stuff i have forgotten.

Hope you have a really great holiday and New Year!

Sybee