ALL Metrics
-
Views
-
Downloads
Get PDF
Get XML
Cite
Export
Track
Study Protocol
Revised

Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol

[version 3; peer review: 3 not approved]
PUBLISHED 23 Apr 2024
Author details Author details
OPEN PEER REVIEW
REVIEWER STATUS

This article is included in the Datta Meghe Institute of Higher Education and Research collection.

Abstract

Platelet indices and the neutrophil:lymphocyte ratio are newly emerging indicators of systemic inflammation in chronic diseases, such as hemoglobinopathies. The objectives of this study include estimation of platelet indices and neutrophil:lymphocyte ratio in sickle cell disease (SCD) patients with and without vaso-occlusive crisis. This cross-sectional study will be conducted in the Department of Medicine at a tertiary care teaching hospital situated in the rural area of Wardha district. The participants will include patients of SCD older than 18 years of age attending the sickle cell clinic. We hypothesize the use of neutrophil:lymphocyte ratio (NLR) and platelet indices as clinically significant predictive markers in acute sickle cell vaso-occlusive crisis.

Keywords

Neutrophil:lymphocyte ratio, Platelet Indices, Sickle cell disease, vaso occlusive crisis

Revised Amendments from Version 2

The reviewer's comments has been addressed thoroughly and vital changes has been made to the article including grammatical corrections.

See the authors' detailed response to the review by Ashwin P. Patel

Introduction

The sickle cell disease (SCD)-causing globin gene mutation can be inherited homozygously or compound heterozygously. The hydrophobic amino acid valine replaces the hydrophilic amino acid glutamic acid in the sixth position to produce haemoglobin S (HbS). HbS polymerizes as a result of deoxygenation, and this polymerization is associated with both increased red cell density and cell dehydration. The disease’s acute (painful episodes, acute chest syndrome) and chronic (avascular necrosis, renal insufficiency) consequences are considered to be caused by the thick, rigid, and sickling red cells, which cause vaso-occlusion and reduced blood flow and are associated with these symptoms.1

In SCD, the complicated interactions between sickled erythrocytes, endothelial cells, leukocytes, platelets, and plasma proteins lead to the vaso-occlusive crisis (VOC). It is generally known that platelets play a part in these interactions. While platelet levels typically rise and are functionally hyperactive in the “steady-state” of sickle cell illness, during a sickle cell crisis, platelet count falls first and then increases, later on, during recovery. Although it is uncommon, acute splenic sequestration, aplastic crisis brought on by parvovirus B19 infection, and fat embolism can all cause thrombocytopenia.2

Vaso-occlusive crises are associated with enhanced platelet reactivity and activation, which raises platelet volume and widens platelet distribution. Moreover, patients with sickle cell anaemia who experienced more crises had higher platelet concentrations than other patients. In contrast to wealthy nations, patients in central India, also referred to as the sickle cell belt, sadly experience more severe symptoms, with vaso-occlusive crisis and stroke being the two main clinical manifestations.3 A number of studies done in this region reflect on the severity of SCD related complications.46

In addition to the pain associated with VOC, there are a number of clinical consequences. Acute chest syndrome, acute bone pain, hepatic and renal involvement, cerebrovascular accident, and multi-organ failure resulting in death can develop in association with SCD which comprises of VOC. All these presentations of SCD will be included in this study.5,6

In several inflammatory and hemolytic illnesses, mean platelet volume serves as a prognostic indicator. Moreover, higher platelet production will result in larger, more reactive platelets with a larger mean platelet size, hence the MPV readings can be utilised to draw conclusions about disorders causing platelet destruction.3

The high cost of hydroxyurea therapy for sickle cell anaemia challenges the significant population that hail from tribal and socioeconomically disadvantaged backgrounds. Also, there hasn’t been much research done on the use of platelet counts as prognostic indicators, which might be very helpful, particularly for a nation like India with a significant population of sickle cell sufferers. As a result, the goal of this study is to evaluate the role of platelet indices in the early identification of patients at a higher risk of crisis. If a positive correlation is found, platelet indices may be used as a required investigation in the evaluation of sickle cell patients to predict the possibility of vaso-occlusive crisis and to start hydroxyurea therapy early for better patient outcomes.3

Objectives

  • To estimate platelet indices in sickle cell disease patients with and without crisis.

  • To estimate neutrophil:lymphocyte ratio in SCD patients with and without crisis.

Protocol

Setting

The study will be conducted in the Department of Medicine, at a tertiary care teaching hospital Acharya Vinoba Bhave Rural hospital situated in the rural area of Wardha district. The overview of the study has been shown in Figure 1.

bdb45c2c-6c9a-4904-8f76-64a9aa3bba5c_figure1.gif

Figure 1. Study flow chart showing the overview of the study.

Study design

This will be a cross-sectional study.

Participants

Cases of sickle cell disease more than 18 years attending Sickle cell clinic at a tertiary care teaching hospital Acharya Vinoba Bhave Rural hospital situated in the rural area of Wardha district.

Inclusion criteria

  • Known patients of SCD older than 18 years of age

  • Newly detected cases of SCD older than 18 years of age

Exclusion criteria

  • Age - less than 18 years

  • Patients with other causes of thrombocytopenia

  • Chronic liver disease and chronic kidney disease

  • Patients with hereditary disorders of large platelets

  • Patients on anticoagulants

  • Patients on antiplatelet agents

Sample size

Daniel et al. formula for sample size:

n=Z22XPX1Pd2

where, Z22 is the level of significance at 5% i.e. 95% confidence interval = 1.96

P=prevalence of sickle cell anemia=4.6%=0.046
D=Desired error of margin=5%=0.05
n=1.962×0.046×10.0460.052=67.43

n = 70 patients needed in each group

Statistical methods: Chi Square test

Student t-test

Software used: SPSS 27.0 Version

Methods

The following data will be collected for all patients with SCD older than 18 years of age.

  • 1. Age of the patient

  • 2. Thorough history – including family history and previous history of crisis, history of chest pain, medication history

  • 3. Assessing pre-existing comorbidities in patient if any:

    • H/O Cancer

    • H/O Chronic Kidney disease

    • H/O Chronic Liver Disease

  • 4. Complete blood count-platelet count and Platelet indices – MPV (mean platelet volume), PDW (platelet distribution width) & PCT (plateletocrit) (blood sample to be collected on day of admission) and differential leucocyte count (neutrophil, lymphocyte, basophils, monocytes and eosinophils)

Lab investigations

Complete blood count with hemoglobin and differential leucocyte count (neutrophil, lymphocytes, eosinophils, basophils and monocytes), platelet count.

Mean platelet volume (fl), Platelet distribution width (%), plateletcrit (%).

Statistical analysis

Data collection will be done on demography, comorbidities, platelet count, MPV, PDW and PCT.

The data will be examined using IBM and SPSS (IBM Corp., Statistics for Windows, version 27.0, Armonk, NY). Continuous variables with a normal distribution are expressed as mean and standard deviation while continuous variables with a non-normal distribution are expressed as median and inter-quartile range. If there is a statistically significant difference between the bivariate samples in independent groups, it will be determined using the unpaired sample t-test and the Chi-square test. For all of the aforementioned statistical techniques, a p-value ≤ 0.05 will be considered as statistically significant.

Dissemination

The report will be published in an indexed journal.

Study status

The study has not yet started.

Discussion

The most often performed laboratory test in patients with sickle cell disease is the complete blood counts. Little is known, however, about the frequency of platelet-count abnormalities, clinical correlation of various forms of sickle cell crises, and the prognostic importance of platelet indices in these critically ill patients in the ICU. This is the main reason behind designing this protocol.3

Common complications include vaso-occlusive crisis and stroke. The main event in pathophysiology of sickle cell crisis is vaso-occlusion with ischemia-reperfusion damage, although the pathophysiology at play is complicated.3 Deoxyhemoglobin S polymerization causes tissue ischemia, which causes both acute and chronic pain. It can also cause damage to any organ in the body including the bones, joints, brain, eyes, liver, kidneys and lungs. SCD activates all cellular components in the blood system.4

Additionally, aberrant red cells and activated platelets may interact, resulting in vaso-occlusion and aggregation. MPV is a simple way to measure platelet activation and function.3,7

In illnesses like idiopathic thrombocytopenic purpura, platelet production and/or destruction are accelerated and MPV, a measure of platelet size, is increased. In individuals with coronary heart disease, higher MPV levels have been found to be an independent risk factor for myocardial infarction, as well as for death or recurrent vascular events following myocardial infarction.4

Studies have linked high NLR with clinical outcomes in cardiovascular disease, malignancies, as well as renal disease in SCD NLR has been proposed as marker of inflammation. The transition from steady state to vaso- occlusive crisis (VOC) is facilitated by leukocytes, which are both elevated and active in SCD. By secreting pro-inflammatory mediators and expressing related adhesion molecules, leukocytes increase the adhesiveness of the endothelial cells in the presence of the proper VOC-producing substances, hence intensifying vaso-occlusion and the pain process.5

Just prior to or during VOC, there is a surge in hematopoietic activity in addition to the shift of the marginating leukocytes to the circulation pool. The significant leukocytosis seen in the VOC group as opposed to the steady state non vaso occlusive SCD patients may be caused by this. The tendency of leukocytes to stick to the postcapillary venular endothelium is increased during leukocytosis, which reduces the vessel lumen. Sickled and unsickled RBCs, platelets and leukocytes all have a greater propensity to cling to the arterial wall and to other cells in these patients, which exacerbates this blockage. Contrarily, in these individuals, leukocytosis might be a type of defence against overwhelming infectious pathogens to which SCD patients are sensitive as a result of functional asplenia brought on by repeated splenic infarction. Patients with functional asplenia are more susceptible to severe infections, especially those caused by encapsulated pathogens such as Streptococcus pneumoniae and Hemophilus influenzae.8

In SCD patients, a high NLR increases the chance of developing VOC and other end-organ damage. In line with John et al., this study aims to discover the relation of NLRs in SCD patients as compared to HbA controls.8 Another study, on the other hand, was unable to show a connection between increased NLR and sickle cell nephropathy, perhaps as a result of the cohort being made up of SCD patients in the steady state.8

In line with earlier studies, a rebound thrombocytosis may have been the cause of the dramatically higher platelet count in the VOC and steady state group. Due to the compensating feedback impact that anaemia has on the synthesis of erythropoietin, rebound thrombocytosis may coexist with hemolytic anemias and the anaemia caused by chronic diseases. The structural similarity between erythropoietin and thrombopoietin is to blame for this. Additionally, hyposplenism in SCD excludes the spleen’s pooling impact on platelets, which greatly contributes to the high platelet count seen in voc state.8

Although there is no statistically significant difference between the steady state and VOC groups in the percentage of haemoglobin S in each red blood cell, it is inversely correlated with HbF levels and inversely proportionate to the degree of polymerization of deoxygenated sickle haemoglobin. However, in all previous studies conducted, NLR was not correlated with HbF.8

The neutrophil and platelet contributions to the chronic inflammatory state in SCD patients are supported by the positive connection between neutrophil to lymphocyte ratio in these individuals. These ratios may serve as reliable, inexpensive and stable indicators of inflammation. When determining the level of inflammation in SCD patients, the use of these simple indicators may be beneficial and superior to using absolute neutrophil count or platelet count alone.8

This is due to the fact that these ratios are a little bit more stable and are less susceptible to the physiological, pathological or stressful circumstances that are known to affect platelet and neutrophil counts.8

Ethical considerations

IEC approval was obtained from Datta Meghe Institute of Medical Sciences and research (Reference number: DMIMS (DU)/IEC/2022/1084).

Written informed consent will be obtained from the patients and their relatives/parents in case of minor patients.

Comments on this article Comments (0)

Version 3
VERSION 3 PUBLISHED 06 Sep 2023
Comment
Author details Author details
Competing interests
Grant information
Copyright
Download
 
Export To
metrics
Views Downloads
F1000Research - -
PubMed Central
Data from PMC are received and updated monthly.
- -
Citations
CITE
how to cite this article
Khurana K and Mahajan S. Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol [version 3; peer review: 3 not approved]. F1000Research 2024, 12:1111 (https://doi.org/10.12688/f1000research.136668.3)
NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.
track
receive updates on this article
Track an article to receive email alerts on any updates to this article.

Open Peer Review

Current Reviewer Status: ?
Key to Reviewer Statuses VIEW
ApprovedThe paper is scientifically sound in its current form and only minor, if any, improvements are suggested
Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
Not approvedFundamental flaws in the paper seriously undermine the findings and conclusions
Version 3
VERSION 3
PUBLISHED 23 Apr 2024
Revised
Views
1
Cite
Reviewer Report 20 Jan 2025
Chisom Adaobi Nri-ezedi, Nnamdi Azikiwe University, Awka, Anambra, Nigeria 
Not Approved
VIEWS 1
Question 1: Is the rationale for, and objectives of, the study clearly described?
Answer: Partly
Comments:
The rationale for the study is vaguely described and does not align well with the research title. While the study aims ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Nri-ezedi CA. Reviewer Report For: Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol [version 3; peer review: 3 not approved]. F1000Research 2024, 12:1111 (https://doi.org/10.5256/f1000research.165308.r360270)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
Views
0
Cite
Reviewer Report 21 Nov 2024
Lewis Hsu, University of Illinois at Chicago, Chicago, Illinois, USA 
Not Approved
VIEWS 0
Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol
Khurana K and Mahajan S

This study protocol has the admirable goal of finding information in the ordinary ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Hsu L. Reviewer Report For: Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol [version 3; peer review: 3 not approved]. F1000Research 2024, 12:1111 (https://doi.org/10.5256/f1000research.165308.r337142)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
Views
5
Cite
Reviewer Report 12 Jun 2024
Ashwin P. Patel, Emory University, Atlanta, USA 
Not Approved
VIEWS 5
I thank authors for editing their protocol. However, some of the previous concerns are not addressed and the protocol contains some ambiguous or contradictory statements. Overall, the protocol fails to convey the clear picture about the proposed research.
    ... Continue reading
    CITE
    CITE
    HOW TO CITE THIS REPORT
    Patel AP. Reviewer Report For: Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol [version 3; peer review: 3 not approved]. F1000Research 2024, 12:1111 (https://doi.org/10.5256/f1000research.165308.r268986)
    NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
    Version 2
    VERSION 2
    PUBLISHED 05 Mar 2024
    Revised
    Views
    8
    Cite
    Reviewer Report 28 Mar 2024
    Ashwin P. Patel, Emory University, Atlanta, USA 
    Approved with Reservations
    VIEWS 8
    I thank the authors for editing their protocol as per my suggestions. Clarification of some aspects of the revised version will be helpful:
    (1) How is acute sickle cell vaso-occlusive crisis defined? Patients usually receive treatment from a local ... Continue reading
    CITE
    CITE
    HOW TO CITE THIS REPORT
    Patel AP. Reviewer Report For: Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol [version 3; peer review: 3 not approved]. F1000Research 2024, 12:1111 (https://doi.org/10.5256/f1000research.163296.r252862)
    NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
    • Author Response 04 Jun 2024
      Kashish Khurana, Medicine, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India
      04 Jun 2024
      Author Response
      The reviewer's comments has been addressed thoroughly and vital changes has been made to the article including grammatical corrections.
      1.) Definition of vaso occlusive crisis has been added to the ... Continue reading
    COMMENTS ON THIS REPORT
    • Author Response 04 Jun 2024
      Kashish Khurana, Medicine, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India
      04 Jun 2024
      Author Response
      The reviewer's comments has been addressed thoroughly and vital changes has been made to the article including grammatical corrections.
      1.) Definition of vaso occlusive crisis has been added to the ... Continue reading
    Version 1
    VERSION 1
    PUBLISHED 06 Sep 2023
    Views
    16
    Cite
    Reviewer Report 24 Jan 2024
    Ashwin P. Patel, Emory University, Atlanta, USA 
    Approved with Reservations
    VIEWS 16
    The authors should address the following queries to increase the clarity of the proposed research.
    (1) The inclusion and exclusion criteria mentioned in the draft and Figure 1 are different. For example, the figure mentions age < 18 years ... Continue reading
    CITE
    CITE
    HOW TO CITE THIS REPORT
    Patel AP. Reviewer Report For: Platelet indices and neutrophil:lymphocyte ratio as a predictive tool in acute sickle cell vaso-occlusive crisis: A study protocol [version 3; peer review: 3 not approved]. F1000Research 2024, 12:1111 (https://doi.org/10.5256/f1000research.149812.r232202)
    NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
    • Author Response 13 Apr 2024
      Kashish Khurana, Medicine, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India
      13 Apr 2024
      Author Response
      1) In the age criteria mentioned in the article, there is a typing mistake and the age criteria in exclusion criteria should be <18 years and not <14 years. It ... Continue reading
    COMMENTS ON THIS REPORT
    • Author Response 13 Apr 2024
      Kashish Khurana, Medicine, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India
      13 Apr 2024
      Author Response
      1) In the age criteria mentioned in the article, there is a typing mistake and the age criteria in exclusion criteria should be <18 years and not <14 years. It ... Continue reading

    Comments on this article Comments (0)

    Version 3
    VERSION 3 PUBLISHED 06 Sep 2023
    Comment
    Alongside their report, reviewers assign a status to the article:
    Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested
    Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
    Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions
    Sign In
    If you've forgotten your password, please enter your email address below and we'll send you instructions on how to reset your password.

    The email address should be the one you originally registered with F1000.

    Email address not valid, please try again

    You registered with F1000 via Google, so we cannot reset your password.

    To sign in, please click here.

    If you still need help with your Google account password, please click here.

    You registered with F1000 via Facebook, so we cannot reset your password.

    To sign in, please click here.

    If you still need help with your Facebook account password, please click here.

    Code not correct, please try again
    Email us for further assistance.
    Server error, please try again.