Prenatal vitamin D3 supplementation suppresses LL-37 peptide

Suikerbuik

Member
Joined
Jan 25, 2014
Messages
700
There's written a lot on vitamin D and as it involves nutrition (and ultimately personal health) I think it's definitely worth posting here. This is another interesting EX VIVO study that doesn't support the view of targeting 40-50 ng/ml and covers some of the discussion points we had (see some quotes form the full-text below).

ABSTRACT
Prenatal vitamin D3 supplementation suppresses LL-37 peptide expression in ex vivo activated neonatal macrophages but not their killing capacity
Vitamin D has regulatory effects on innate immunity. In the present study, we aimed to assess the effect of prenatal vitamin D3 (vitD3) supplementation on neonatal innate immunity in a randomised, placebo-controlled trial by evaluating cathelicidin (LL-37) expression and the killing capacity of macrophages. Healthy pregnant women (n 129) attending a clinic in Dhaka were randomised to receive either a weekly oral dose of 0·875 mg vitD3 or placebo starting from 26 weeks of gestation up to delivery. Serum, plasma and monocyte-derived macrophages (MDM) were obtained from the cord blood. 25-Hydroxyvitamin D (25(OH)D) concentration was measured in serum. MDM were stimulated with or without Toll-like-receptor 4 ligand (TLR4L). Innate immune function was assessed by measuring LL-37 peptide levels in the culture supernatant of MDM by ELISA, LL-37 transcript levels by quantitative PCR, and ex vivo bactericidal capacity of MDM. vitD3 supplementation did not increase LL-37 peptide levels in plasma or in the extracellular fluid of macrophages with or without TLR4L induction. However, stimulated intracellular LL-37 expression (ratio of stimulated:unstimulated MDM) was significantly reduced in the vitamin D group v. placebo (P= 0·02). Multivariate-adjusted analyses showed that intracellular LL-37 peptide concentration from stimulated MDM was inversely associated with 25(OH)D concentration in serum (P= 0·03). TLR4L stimulation increased the bactericidal capacity of MDM compared with the unstimulated ones (P= 0·01); however, there was no difference in killing capacity between the two groups. A weekly dose of 0·875 mg vitD3 to healthy pregnant women suppressed the intracellular LL-37 peptide stores of activated macrophages, but did not significantly affect the ex vivo bactericidal capacity of cord blood MDM.
http://www.ncbi.nlm.nih.gov/pubmed/25089537

0.875 mg = ~34000 i.u. (per week)

I'll quote some statements or additional information on the subject from the full text:

All major immune cells, including T cells, B cells, neutrophils and antigen-presenting cells (monocytes, macrophages and dendritic cells), express the vitamin D receptor. Macrophages, the front-line responders to microbial infection, sense pathogen-associatedmolecularpatterns by utilising pattern recognition receptors such as Toll-like receptors (TLR). TLR triggering induces Cytochrome p450 27B1 (CYP27B1), a vitamin D-activating enzyme that converts 25-hydroxyvitamin D (25(OH)D) into the active form (1,25-dihydroxyvitamin D; 1,25(OH)2D), and vitamin D receptor expression(3). The 1,25(OH)2D–vitamin D receptor complex directly induces cathelicidin antimicrobial peptide (CAMP; gene encoding LL-37) transcription and corresponding increases in the expression of the antimicrobial peptide, LL-37(4).

Cord serum 25(OH)D was not significantly associated with LL-37 concentration in the unstimulated ICF, or fold difference in intracellular LL-37 concentration (Table 2). However, 25(OH)D in cord serum was inversely related to TLR4L-induced intracellular LL-37 in both unadjusted bivariate and adjusted multivariate analyses (Table 2). For every 10 nmol/l increase in cord serum 25(OH)D concentration, TLR4L-induced intracellular LL-37 concentration decreased by 0·05 ng/ml (P¼0·03).

Upon stratifying the participants into four groups based on serum 25(OH)D concentrations, LL-37 peptide concentrations in the ICF of TLR4L-stimulated MDM were significantly reduced in the high (>76nmol/l), moderate (50–75 nmol/l) and low (30–49nmol/l) groups (P¼0·005, P¼0·035 and P¼0·029, respectively) compared with the very low (<30nmol/l) group (Fig. 1(a)).

the present study showed that prenatal vitD3 supplementation did not yield significant benefits in terms of indicators of innate immune function. In fact, we observed that vitD3 supplementation may have slightly suppressed intracellular LL-37 peptide synthesis in activated cord blood macrophages, suggesting a potential immunosuppressive effect. Furthermore, we did not find any significant influence of vitD3 supplementation on the bactericidal capacity of macrophages with or without TLR4L activation. In vitro studies using primary monocytes/macrophages cultured in 25(OH)D-sufficient sera or culture media containing the active form of vitamin D (1,25(OH)2D3) have shown that vitamin D mediates the induction of cathelicidin (LL-37) and b-defensin 4 transcripts in macrophages(3,24,25). In vitro addition of 1,25(OH)2D3 or 25(OH)D3 in cord blood cultures also showed increased TLR-independent or -dependent cathelicidin mRNA expression(23). In the present study, in vivo vitD3 supplementation did not affect LL-37 mRNA levels in neonatal macrophages with or without activation. The main difference between the other reports and the present study is that here vitD3 was given orally to pregnant women and autologous plasma was used instead of adding 1,25(OH)2D3 or 25(OH)D3 exogenously in the macrophage culture.

The present results thus collectively suggest that in vitro findings related to the effect of vitamin D metabolites on LL-37 expression may not be consistent and may not predict the in vivo effects of vitD3 supplementation,

In the abovementioned previous study involving healthy adult volunteers, we found that vitD3 supplementation significantly increased the antibacterial killing activity of macrophages in the absence of any increase in the levels of LL-37 peptide or mRNA expression(31). However, there are some key differences between our two studies. In our previous study(31), vitD3 supplementation was administered for only 4 d (even though the daily dose was the same), while in the present study, supplementation was given for about 3 months(19). The previous study involved healthy adults, while the participants of the present study were healthy pregnant women, and immune function was studied in cord blood, not in maternal specimens.

So another decent study in which data supports against the use of suppletion, at least shows no single benefit from the use of vitamin D. And provides support to the view of healthy levels to be between 20- 30 ng/ml instead of levels >30 ng/ml.
 
Joined
Nov 26, 2013
Messages
7,370
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

I still don't get why we should spend money on yet another supplement when fifteen minutes of sun is all that is needed.
 

Wilfrid

Member
Joined
Nov 26, 2012
Messages
723
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

@Suikerbuik,

Thanks so much for all those precious info about vitamin D supp.

@Such_Saturation,

:1
 

BingDing

Member
Joined
Nov 20, 2012
Messages
976
Location
Tennessee, USA
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

Suikerbuik, you have been pretty relentless in promoting your views about Vit D3 and blood tests for 25-OH D3, and I am getting a little tired of it. Nothing that Ray has said or written supports your view, AFAIK, and he has written about Vit D a lot.

Maybe you could email Ray and see if the studies you have posted change his mind.

For me, studies about prenatal influence on an obscure metabolic marker are pretty abstract, and don't inform about the issue of optimum Vit D levels or supplementation.
 
OP
Suikerbuik

Suikerbuik

Member
Joined
Jan 25, 2014
Messages
700
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

Hi BingDing,

Hehe, I can completely understand. I had planned to shut up after all the discussion some months ago. But after your comment:
What is most disappointing is that D3 is probably one of the easiest nutrients to get right.
I could not resist answering and feedback some statements that you made. I wanted to know why someone would think like that, what formed his conclusions or maybe you knew something I didn't know yet.

There were also some open ends in the discussions before, because of not well performed studies (e.g., in vitro, the use of 1,25D, treatment duration, dose usage). All these aspects are involved in this study and see what they come up with.

LL-37 is an incredible important peptide, called an antimicrobial peptide. These peptides are our own antibiotics. This peptide is expressed in a variety of immune cells and epithelial cells (skin, gastrointestinal tract, respiratory tract). It plays a key role in innate immunity (non-specific immunity). What is more interesting, is that prenatal babies have to rely on this type of immunity for defense (hence innate), they haven't yet a good functional adaptive immune system this evolves during life. In this stage mother's milk is providing the antibodies. What next is that the "activated" VDR complex is a strong inducer of this peptide. So what you would think is that 25D suppletion increases LL-37, but this is not the case. And with levels over 75 nmol/L also LL-37 transcription recedes. The expressions of this pepitde is altered in certain conditions. Ofcourse this is not in adults human, but still tells us pretty much in what kind of direction we have to look.

I don't have Peat's email but yes, I would highly appreaciate his view on this subject after these recent studies.

But no worries anymore. I am not planning to make any new thread, or place more comments on the subject. This study and all the other previous comments elsewhere should provide enough information to think about the subject yourself, and not someone involved in the vitamin D bussines.
Only, if I somehow have been in contact with Dr. Peat, I'll post how he thinks about all this.
 

LucyL

Member
Joined
Oct 21, 2013
Messages
1,244
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

Suppressing (regulating?) LL-37 peptide isn't defacto a bad thing. To know whether the suppression of the LL-37 peptide in pregnant women in Bangladesh has a negative or positive effect, would require followup on the long term health of the babies. Dhaka is a very densely populated city in a very densely populated country, with a lot of smog, pollution and other stressors that go hand in hand with crowded living conditions. I would like to know what autoimmune disease rates are like in that city. Diabetes rates in Bangladash for instance, are sky high compared to other (UN designated) "least developed" countries.

The study said the women were healthy, but what does that really mean?


]Little peptide, big effects: the role of LL-37 in inflammation and autoimmune disease.
Kahlenberg JM1, Kaplan MJ.
Author information
Abstract

The innate immune system utilizes many approaches for defense against invading microorganisms, including complement-mediated lysis, engulfment, formation of neutrophil extracellular traps, and release of antimicrobial peptides. Although classically thought to be driven by adaptive immunity, the development of autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus is increasingly associated with dysregulated innate immune pathways. An emerging theme within this literature is the contribution of antimicrobial peptides to the development of autoimmune disorders. This is best exemplified in atopic dermatitis and psoriasis where the defensins and the single human cathelicidin, LL-37, may contribute to disease. Furthermore, in the past few years, a role for LL-37 has emerged in the pathogenesis of systemic lupus erythematosus, rheumatoid arthritis, atherosclerosis, and possibly other diseases. In this review, we discuss the role of LL-37 and its murine ortholog, mCRAMP, in the modulation of immune and inflammatory pathways and their effects on autoimmune and inflammatory diseases.
 
OP
Suikerbuik

Suikerbuik

Member
Joined
Jan 25, 2014
Messages
700
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

You have the full text for me? To me this just looks like a random paper that puts LL-37 in bad lights. I am not surprised by that, go look further and you will find more. Try take a look at LL-37 and cancer for example, or whatever. We have seen it before even recently on this forum, two opposite conclusions from the same data. I want to see the data myself, and know what kind of hypothesis they had initially.

If you do so, also take a look what an active vitamin D receptor does for cancer. And how LL-37 seems to be regulated – mainly by an “activated” vitamin D receptor complex.

The fact that LL-37 in some cases is upregulated, doesn’t mean that LL-37 is the origin of health problems. Ofcourse it could play a role in pathogenesis, it still involves the immune system. But are there studies that show patient outcomes when LL-37 is suppressed by some medicine? Only then we can make conclusions whether this is good or not. I mean they can find a correlation between pathogenesis and some metabolite but perhaps suppressing that metabolite would even be more deletorious? (could very well be the case as seen in TNF-a blockers). You will find a negative for anything ;)

I find “auto-immune” disorders scary to talk about to be honest. I don’t think the body mounts a response without a reason, or it has to involve molecular mimicry (then there’s still a reason though). Science just doesn't know yet what the exactly the underlying reason is in most cases and this will differ among individuals. (I am aware of Peat's articles for example, but these do not provide all the answers, though very important information for people suffering). Despite our poor knowlegde on causes, our understanding of "pathological" processes increases. We like to chase the tail and invent new medicines to make more money on the backs of sick people, I guess.

I disagree about a follow up of the babies. We can “measure” the health of those babies and LL-37 and try find a correlation, but what about the thousands of other metabolites (and environment altering these metabolites) that we don’t track and may have a far more significant impact on health outcome? In the end LL-37 is just another metabolite, but a metabolite that says much about VDR activation. This is what is so interesting, because as expected by current (in vitro mainly) research on vitamin D, you are told that 25D is the storage hormone, will not interfere with vitamin D receptor activation and will likely only increase it. Turns out no to be so, at least not in theses cases.

Health is something incredible difficult to describe. According to some numbers/ markers half of the western population is diseased. You are right about all the environmental aspects. But does that matter for vitamin D and LL-37 expression status? Is there any evidence that any of those environmental aspects can affect LL-37 status directly?
It would certainly be interesting so this the same study being done in lets say some african tribe.
 

LucyL

Member
Joined
Oct 21, 2013
Messages
1,244
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

Suikerbuik said:
You have the full text for me?

Sorry, I can't find the full text online anywhere.

Suikerbuik said:
The fact that LL-37 in some cases is upregulated, doesn’t mean that LL-37 is the origin of health problems. Ofcourse it could play a role in pathogenesis, it still involves the immune system. But are there studies that show patient outcomes when LL-37 is suppressed by some medicine? Only then we can make conclusions whether this is good or not. I mean they can find a correlation between pathogenesis and some metabolite but perhaps suppressing that metabolite would even be more deletorious? (could very well be the case as seen in TNF-a blockers). You will find a negative for anything ;)

MD Anderson researchers have reportedly demonstrated that
LL37 activates the dendritic cells by binding to the self-DNA to form a structure that allows it into the dendritic cells, as if it were an invading microbe.

Of course they are looking for drugs to block LL37, which would probably be absolutely disastrous. Anyway, why invent something new if Vit D already regulates it well?

Suikerbuik said:
I disagree about a follow up of the babies. We can “measure” the health of those babies and LL-37 and try find a correlation, but what about the thousands of other metabolites (and environment altering these metabolites) that we don’t track and may have a far more significant impact on health outcome?

Yes, but given the known relation (whatever it really is) between LL37 and autoimmune disorders, you almost have to see if suppressing it during pregnancy has any correlation to the development of autoimmune disorders later in life.Curiosity demands it :D


Suikerbuik said:
It would certainly be interesting so this the same study being done in lets say some african tribe.

It is already known that Vit D has different effects dependent on race. Effect of Vitamin D Supplementation . . . To be applicable to us (well, me anyway ;-) ) it would have to be done on a standard first world Caucasian population. Another variable.
 
OP
Suikerbuik

Suikerbuik

Member
Joined
Jan 25, 2014
Messages
700
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

Hi LucyL,

thank you I appreciate your commets !

This can be an ongoing discussion, neither you nor I is wrong I think. This is because of all the variables and from which glasses you look at things. But I’ll try to comment shortly.

Yes, but given the known relation (whatever it really is) between LL37 and autoimmune disorders
LL-37 is just one of a multitude of antimicrobial peptides with little overall correlation when compared to ALL autoimmune or infectious diseases in general. But a role for the activated vitamin D receptor complex in regulating LL-37 is well known.

But in the case of autoimmune disease, LL-37 remains persistently produced, well beyond the temporary jolt needed to dampen infection and promote healing.
So because it's persistently upregulated means that LL-37 should be a target? LL-37 doesn’t transcribe itself as far as I know. I think it’s the same with like tumors that induce autoimmunity. The auto immunty disappears when the tumor is targeted/ gone. We have far from discovered everything. A recent study even found viable microbiota in woman’s breast tissue. see here

Lately we were talking about boundaries, and although boundaries do exist these do not mean that they are impermeable or permanent. I don't see a distinct boundary between most tissue. And a boundary is only a boundary for something if the resistance can’t be overcome. What if it turns out if every tissue contains microbiota? Science would be upside down, but until this is disproved I'd consider this as an option.

Thanks for pointing towards this paper I have seen it before. And although there are certainly differences between the races. A tendency between vitamin D suppletion and LL-37 expression is absolutely observed (see table 1). But I would indeed not attach to the numbers they find.
 

BingDing

Member
Joined
Nov 20, 2012
Messages
976
Location
Tennessee, USA
Re: Prenatal vitamin D3 supplementation suppresses LL-37 pep

Suikerbuik, I want to apologize if my remarks above were a little blunt. I had been reading your posts as advice for people to follow, and was frustrated because I couldn't make sense of it. I now understand that might not have been the context you intended.

I agree that discussing and explaining studies and ideas is completely appropriate here, and I welcome more of it.

BD
 

Similar threads

Back
Top Bottom